U.S. patent application number 15/242330 was filed with the patent office on 2017-06-15 for imidamide sphingosine kinase inhibitors.
The applicant listed for this patent is University of Virginia Patent Foundation. Invention is credited to Andrew Kennedy, Yugesh Kharel, Kevin R. Lynch, Timothy L. Macdonald, Thomas P. Mathews.
Application Number | 20170166525 15/242330 |
Document ID | / |
Family ID | 43586552 |
Filed Date | 2017-06-15 |
United States Patent
Application |
20170166525 |
Kind Code |
A1 |
Lynch; Kevin R. ; et
al. |
June 15, 2017 |
IMIDAMIDE SPHINGOSINE KINASE INHIBITORS
Abstract
Imidamide (amidine) analogs that can inhibit the activity of
sphingosine kinase 1 and sphingosine kinase 2 (SphK1 & SphK2)
are provided. The compounds can prevent angiogenesis in tumors.
Inventors: |
Lynch; Kevin R.;
(Charlottesville, VA) ; Macdonald; Timothy L.;
(Charlottesville, VA) ; Mathews; Thomas P.;
(Charlottesville, VA) ; Kennedy; Andrew;
(Charlottesville, VA) ; Kharel; Yugesh;
(Charlottesville, VA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
University of Virginia Patent Foundation |
Charlottesville |
VA |
US |
|
|
Family ID: |
43586552 |
Appl. No.: |
15/242330 |
Filed: |
August 19, 2016 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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13390207 |
Apr 27, 2012 |
9421177 |
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PCT/US10/45660 |
Aug 16, 2010 |
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15242330 |
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61233963 |
Aug 14, 2009 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 31/40 20130101;
A61P 37/00 20180101; A61P 43/00 20180101; A61P 27/02 20180101; C07C
2601/02 20170501; A61P 11/06 20180101; A61P 29/00 20180101; C07D
207/09 20130101; A61P 19/02 20180101; A61P 25/00 20180101; C07C
2601/08 20170501; C07C 2601/14 20170501; A61P 3/10 20180101; A61P
9/00 20180101; C07C 2603/74 20170501; A61P 11/00 20180101; A61P
35/00 20180101; A61P 1/00 20180101; A61K 31/155 20130101; A61P 1/16
20180101; A61P 13/12 20180101; C07C 257/16 20130101 |
International
Class: |
C07D 207/09 20060101
C07D207/09; C07C 257/16 20060101 C07C257/16 |
Goverment Interests
GOVERNMENT FUNDING
[0002] This invention was made in part with United States
Government support under Grant No. RO1 GM 067958 awarded by the
National Institutes of Health. The United States Government has
certain rights in the invention.
Claims
1. A compound of formula IA: ##STR00086## wherein R.sup.1 is
hydrogen or OH; R.sup.2 and R.sup.1 are independently hydrogen,
(C.sub.1-C.sub.4)alkyl; or halo(C.sub.1-C.sub.3)alkyl; R.sup.4 is
NH.sub.2 or (C.sub.1-C.sub.4)alkyl; R.sup.5 is hydrogen or
(C.sub.1-C.sub.4)alkyl; X and Y are independently CR.sup.6R.sup.7,
NR.sup.8, O, S or C.dbd.O (carbonyl); R.sup.6 and R.sup.7 are
independently H or (C.sub.1-C.sub.7)alkyl; R.sup.8 is H or
(C.sub.1-C.sub.7)alkyl; wherein R.sup.4 and one of R.sup.5,
R.sup.6, R.sup.7, or R.sup.8 together with the atoms to which they
are attached form a ring, optionally including one or more
heteroatoms; wherein the heteroatoms are O, S, N or NR.sup.10;
R.sup.10 is hydrogen, or (C.sub.1-C.sub.7)alkyl; Z is
(C.sub.6-C.sub.20)alkyl, (C.sub.6-C.sub.20)alkenyl,
(C.sub.6-C.sub.20)alkynyl, (C.sub.6-C.sub.20)alkoxy,
(C.sub.6-C.sub.26)alkoxyalkyl, (C.sub.3-C.sub.12)cycloalkyl,
(C.sub.6-C.sub.10)aryl, (C.sub.12-C.sub.20)alkylaryl,
(C.sub.12-C.sub.20)alkoxylaryl (C.sub.1-C.sub.30)arylalkyl,
(C.sub.2-C.sub.10)heterocyclic, (C.sub.4-C.sub.10)heteroaryl, or
(C.sub.4-C.sub.10)heteroaryl(C.sub.1-C.sub.20)alkyl; and R.sup.10
is hydrogen, or (C.sub.1-C.sub.7)alkyl; wherein the Z groups are
optionally substituted with 1, 2, 3, 4, 5, 6, or 7 substituents;
where the substituents are halo, halo(C.sub.1-C.sub.10)alkyl,
cyano, --NR.sup.aR.sup.b, (C.sub.1-C.sub.20)alkoxy,
(C.sub.2-C.sub.26)-alkoxyalkyl, (C.sub.3-C.sub.12)cycloalkyl,
(C.sub.6-C.sub.10)aryl, (C.sub.7-C.sub.30)alkylaryl,
(C.sub.2-C.sub.10)heterocyclic, or (C.sub.4-C.sub.10)-heteroaryl;
wherein one or more of the carbon atoms in the Z alkyl groups can
be independently replaced with non-peroxide oxygen, sulfur or
NR.sup.c; where each R.sup.a, R.sup.b, or R.sup.c is independently
hydrogen or (C.sub.1-C.sub.7)alkyl; or a pharmaceutically
acceptable salt or ester thereof.
2. The compound of claim 1, wherein R.sup.2 and R.sup.3 are
hydrogen.
3. The compound of claim 2, wherein R.sup.1 is hydrogen.
4. The compound of claim 2, wherein R.sup.1 is --OH.
5. The compound of claim 1, wherein R.sup.4 and R.sup.5 together
form a cyclopropyl, cyclobutyl or cyclopentyl ring.
6. The compound of claim 5, wherein Z is (C.sub.10-C.sub.18)alkyl,
(C.sub.10-C.sub.18)alkenyl, (C.sub.10-C.sub.18)alkynyl,
(C.sub.10-C.sub.18)alkoxy, (C.sub.12-C.sub.22)alkoxyalkyl,
(C.sub.3-C.sub.10)cycloalkyl, (C.sub.6)aryl,
(C.sub.7-C.sub.18)alkylaryl, (C.sub.7-C.sub.18)alkoxyaryl, or
(C.sub.7-C.sub.18)arylalky.
7. The compound of claim 6, wherein the Z groups are substituted
with 1, 2, or 3 substituents; where the substituents are
(C.sub.3-C.sub.10)cycloalkyl, (C.sub.6)aryl, or
(C.sub.7-C.sub.12)alkylaryl.
8. The compound of claim 1 wherein one of X and Y is NH and the
other is C.dbd.O.
9. The compound of claim 1, wherein Z is
(C.sub.5-C.sub.12)alkyl-4-phenyl.
10. The compound of claim 1, wherein Z is
(C.sub.5-C.sub.12)alkyl-4-phenyl, substituted with
(C.sub.5-C.sub.10)cycloalkyl-O--, phenyl-CH.sub.2--.
11. The compound of claim 1 wherein cycloalkyl is cyclopentyl,
cyclohexyl, or adamantyl.
12. The compound of claim 1, wherein X is NR, R.sup.5 is hydrogen
and R.sup.4 and R.sup.8 form a five membered heterocyclic ring.
13. The compound of claim 12, wherein Z is ##STR00087##
##STR00088##
14. The compound of claim 1 having the formula: ##STR00089##
##STR00090## or a pharmaceutically acceptable salt thereof.
15.-20. (canceled)
21. A method for prevention or treatment of a pathological
condition or symptom in a mammal, wherein inhibition of the SphK1
enzyme or SphK2 enzyme is implicated and such inhibition is
desired, comprising administering to said mammal an effective
amount of a compound of claim 1.
22. (canceled)
23. The method of claim 21, wherein the pathological condition
comprises a vascular injury, fibrosis, sepsis, asthma, or an
autoimmune disease.
24. The method of claim 23, wherein the autoimmune disease is
uveitis, type I diabetes, rheumatoid arthritis, inflammatory bowel
diseases, or multiple sclerosis.
25. The method of claim 23, wherein the autoimmune disease is
multiple sclerosis.
26. The method of claim 21, wherein the pathological condition
comprises asthma.
27.-28. (canceled)
29. The method of claim 23, wherein fibrosis includes pulmonary
renal or hepatic fibroses.
30.-33. (canceled)
Description
PRIORITY
[0001] This application claims priority from U.S. Provisional
Application No. 61/233,963, filed Aug. 14, 2009, the disclosure of
which is incorporated by reference.
BACKGROUND
[0003] Sphingosine 1-phosphate (S1P) is a lysophospholipid mediator
that evokes a variety of cellular responses by stimulation of five
members of the endothelial cell differentiation gene (EDG) receptor
family. The EDG receptors are G-protein coupled receptors (GPCRs)
and on stimulation propagate second messenger signals via
activation of heterotrimeric G-protein alpha (G.sub..alpha.)
subunits and beta-gamma (G.sub..beta..gamma.) dimers. Ultimately,
this S1P-driven signaling results in cell survival, increased cell
migration and, often, mitogenesis. The recent development of
agonists targeting S1P receptors has provided insight regarding the
role of this signaling system in physiologic homeostasis. For
example, the immuno-modulator, FTY720 (2-amino-2-[2-(4-octylphenyl)
ethyl] propane 1,3-diol), that following phosphorylation, is an
agonist at 4 of 5 S1P receptors, revealed that enhancing S1P tone
influences lymphocyte trafficking. Further, S1P type 1 receptor
(S1P1) antagonists cause leakage of the lung capillary endothelium,
which suggests that S1P may be involved in maintaining the
integrity of the endothelial barrier in some tissue beds. S1P is
synthesized by the action of two enzymes, sphingosine kinase types
1 and 2 (SphK1, SphK2), in transferring a phosphate residue from
ATP to sphingosine (the enzymes also catalyze the phosphorylation
of sphinganine to yield sphinganine 1-phosphate (dihydroS1P)).
[0004] Sphingosine 1-phosphate (S1P) has been demonstrated to
induce many cellular processes, including those that result in
platelet aggregation, cell proliferation, cell morphology,
tumor-cell invasion, endothelial cell chemotaxis and angiogenesis.
For these reasons, sphingosine kinases are good targets for
therapeutic applications such as wound healing and tumor growth
inhibition.
[0005] The importance of sphingosine kinase 1 and 2 (SphK1 &
SphK2) in survival and proliferation has also been recognized.
SphK1 & SphK2 catalyze the phosphorylation of the endogenous
lipid D-erythro sphingosine to sphingosine 1-phosphate (S1P). SphK1
& SphK2 are also responsible for the equilibrium between the
anti-apoptotic S1P and its pro-apoptotic metabolic precursor
sphingosine and its precursor, ceramide. Thus, SphK1 & SphK2
have been proposed to be important drug targets. However, only a
small number of compounds have been shown to inhibit the
sphingosine kinases, including DL-threo-dihydrosphingosine,
N,N-dimethylsphingosine and short-chain DL-erythro-sphingosine
analogues. However, these compounds are not suitable as in vivo
inhibitors and cannot address questions concerning SphK mediated
disease states.
[0006] Traditional methods of inhibiting sphingosine kinases have
centered on targeting the ATP binding site of the kinase, a
strategy that has enjoyed moderate success. However, such methods
suffer from limited of selectivity across a wide array of kinases.
Additionally, the sequence of the ATP binding domain of SphK1 &
SphK2 is conserved across a number of diacylglycerol (DAG) kinase
family members, rendering the traditional strategy problematic.
[0007] Currently, there is a need for novel, potent, and selective
agents that inhibit the substrate-binding domain of the sphingosine
kinases (e.g., human SphK1 and SphK2) that have enhanced potency,
selectivity, and oral bioavailability. In addition, there is a need
in the art for identification of, as well as the synthesis and use
of such compounds. The present invention satisfies these needs.
SUMMARY
[0008] The present invention provides, in one aspect, compounds
that inhibit sphingosine kinase 1 and sphingosine kinase 2 (SphK1
& SphK2) enzymes. Accordingly, there is provided compounds of
Formula IA:
##STR00001##
[0009] wherein: R.sup.1 is hydrogen or OH; R.sup.2 and R.sup.3 are
independently hydrogen, (C.sub.1-C.sub.4)alkyl; or
halo(C.sub.1-C.sub.3)alkyl;
[0010] R.sup.4 is NH.sub.2 or (C.sub.1-C.sub.4)alkyl;
[0011] R.sup.5 is hydrogen or (C.sub.1-C.sub.4)alkyl;
[0012] X and Y are independently CR.sup.6R.sup.7, NR.sup.8, O, S or
C.dbd.O (carbonyl); R.sup.6 and R.sup.7 are independently H or
(C.sub.1-C.sub.7)alkyl; R.sup.8 is H or (C.sub.1-C.sub.7)alkyl;
[0013] X and Y are independently CR.sup.6R.sup.7, NR.sup.8, O, S or
C.dbd.O (carbonyl);
[0014] R.sup.6 and R.sup.7 are independently H or
(C.sub.1-C.sub.7)alkyl; R.sup.8 is H or (C.sub.1-C.sub.7)alkyl;
[0015] wherein R.sup.4 and one of R.sup.5, R.sup.6, R.sup.7, or
R.sup.8 together with the atoms to which they are attached form a
ring, optionally including one or more heteroatoms; wherein the
heteroatoms are O, S, N or NR.sup.10; where R.sup.10 is hydrogen,
or (C.sub.1-C.sub.7)alkyl;
[0016] Z is (C.sub.6-C.sub.20)alkyl, (C.sub.6-C.sub.20)alkenyl,
(C.sub.6-C.sub.20)alkynyl, (C.sub.6-C.sub.20)alkoxy,
(C.sub.6-C.sub.26)alkoxyalkyl, (C.sub.6-C.sub.12)cycloalkyl,
(C.sub.6-C.sub.10)aryl, (C.sub.12-C.sub.20)alkylaryl,
(C.sub.12-C.sub.20)alkoxylaryl (C.sub.7-C.sub.30)arylalkyl,
(C.sub.2-C.sub.10)heterocyclic, (C.sub.4-C.sub.10)heteroaryl, or
(C.sub.4-C.sub.10)heteroaryl(C.sub.1-C.sub.20)alkyl; and R.sup.10
is hydrogen, or (C.sub.1-C.sub.7)alkyl; wherein the Z groups are
optionally substituted with 1, 2, 3, 4, 5, 6, or 7 substituents;
where the substituents are halo, halo(C.sub.1-C.sub.10)alkyl,
cyano, --NR.sup.aR.sup.b, (C.sub.1-C.sub.20)alkoxy,
(C.sub.2-C.sub.26)alkoxyalkyl, (C.sub.3-C.sub.12)cycloalkyl,
(C.sub.6-C.sub.10)aryl, (C.sub.7-C.sub.30)alkylaryl,
(C.sub.2-C.sub.10)heterocyclic, or (C.sub.4-C.sub.10)-heteroaryl;
wherein one or more of the carbon atoms in the Z alkyl groups can
be independently replaced with non-peroxide oxygen, sulfur or
NR.sup.c; where each R.sup.a, R.sup.b, or R.sup.c is independently
H or (C.sub.1-C.sub.7)alkyl. The invention includes
pharmaceutically acceptable salts of the compounds of Formula
IA.
[0017] In another aspect, the disclosed compounds and methods are
directed to SphK enzyme inhibitors that have activity as selective
inhibitors of the SphK1 enzyme or SphK2 enzyme or have activity as
inhibitors of both SphK1 and SphK2 enzymes.
[0018] In another aspect, the present invention provides a method
for inhibiting angiogenesis in a tumor, including contacting
cancerous cells with an effective amount of a compound of Formula
IA, or a pharmaceutically acceptable salt or ester thereof.
[0019] In another aspect, the present invention provides
compositions and methods for the use of SphK inhibitors for the
treatment of neoplastic disease. In one aspect, this treatment is
effected by application of SphK inhibitors that are efficacious by
virtue of their anti-angiogenic properties.
[0020] In another aspect, the present invention provides a method
for preventing or treating diseases that involve excess vascular
growth, e.g. retinal degenerative diseases such as macular
degeneration, comprising contacting the affected area with an
effective amount of the compound of Formula IA. For example the
compound can be injected into the posterior eye in depot form.
[0021] In another aspect, the invention provides a method for
repairing a vascular injury following catheterization, including
contacting the lumen of the affected vessel with an effective
amount of the compound of Formula IA. In another aspect, the
invention includes coating indwelling stents with a compound of
Formula IA.
[0022] In another aspect, the invention provides a method for
parenteral delivery of a compound of formula IA prior to and/or
following intravascular stenting.
[0023] In another aspect, the present invention provides
compositions and methods for the use of SphK inhibitors to prevent
and inhibit vascular restenosis following a vascular injury. For
example, the injury can be due to balloon angioplasty. In another
aspect, the present invention includes a method for treating
subjects to prevent vascular restenosis.
[0024] In another aspect, the present invention provides
compositions and methods for the use of SphK inhibitors to prevent
asthma attacks. In one aspect, the asthma could be due to over
production of cysteinyl leukotrienes. In another aspect, the
present invention includes a method for treating subjects suffering
from asthma.
[0025] In another aspect, the present invention provides a metal
stent coated directly on the surface with of a compound of Formula
IA. The compound of Formula IA is delivered to the stented vessel
wall by direct contact of the stent struts with the vessel
wall.
[0026] In another aspect, the present invention provides a compound
of Formula IA incorporated into a non-degradable polymer or
co-polymer, such as but not restricted to polyethylene-co-vinyl
acetate (PEVA) and poly n-butyl methacrylate, and coating a bare
metal stent with the polymer.
[0027] In another aspect, the present invention provides a compound
of Formula IA is incorporated into a biodegradable polymer or
co-polymer, such as but not limited to poly lactic acid glycolic
acid (PLGA) or phosphorylcholine, and coating a bare metal stent
with the polymer. The compound of Formula IA is delivered to the
stented vessel wall by elution from the biodegradable polymeric
surface on the stent.
[0028] In another aspect, the present invention provides a compound
of Formula IA incorporated into a stent device that contains a
nanoporous surface modification, such as but not limited to a
ceramic, metal or other material coated on the bare metal stent as
a nanoporous surface modification. The compound of Formula IA is
delivered to the stented vessel wall by elution from the nanoporous
surface.
[0029] In another aspect, the present invention provides a
non-metallic biodegradable or non-degradable stent device combined
with a compound of Formula IA. The compound of Formula IA is
delivered to the stented vessel wall by elution from the
non-metallic biodegradable or non-biodegradable stent matrix.
[0030] In another aspect, the invention provides a compound of
Formula IA, a pharmaceutically acceptable salt or ester thereof for
use in medical treatment (for example, treatment of neoplastic
disease).
[0031] In another aspect, the invention provides a compound of
Formula IA, a pharmaceutically acceptable salt or ester thereof for
use in medical treatment for example treatment of fibrotic disease
such as pulmonary fibrosis.
[0032] In another aspect, the invention provides a compound of
Formula IA, a pharmaceutically acceptable salt or ester thereof for
use in medical treatment for example treatment of sepsis.
[0033] In another aspect, the invention provides a compound of
Formula IA, a pharmaceutically acceptable salt or ester thereof for
use in medical treatment for example treatment of autoimmune
diseases such as multiple sclerosis, psoriasis, rheumatoid
arthritis or uveitis.
[0034] In another aspect, the invention provides a method for the
use of a compound of Formula IA or a pharmaceutically acceptable
salt or ester thereof to prepare a medicament for inhibiting tumor
growth, metastasis or tumor angiogenesis. In another aspect, the
invention provides a method for the use of a compound of Formula IA
or a pharmaceutically acceptable salt or ester thereof to prepare a
medicament for preventing or treating a vascular injury, fibrosis,
sepsis, asthma, or an autoimmune disease in a mammalian species
(for example, a human).
[0035] The present invention also includes pharmaceutical
compositions including the disclosed compounds. More particularly,
such compounds can be formulated as pharmaceutical compositions
using standard pharmaceutically acceptable carriers, fillers,
solubilizing agents and stabilizers known to those skilled in the
art. For example, a pharmaceutical composition including a
disclosed compound, analog, derivative, or modification thereof, as
described herein, is used to administer the appropriate compound to
a subject.
[0036] In another aspect, the invention provides for the use of a
compound of Formula IA or a pharmaceutically acceptable salt or
ester thereof to prepare a medicament for repairing a vascular
injury following catheterization. In another aspect, the invention
includes use of a compound of Formula IA for coating indwelling
stents.
[0037] In another aspect, the invention provides novel
intermediates and processes disclosed herein that are useful for
preparing compounds of Formula IA, including the generic and
specific intermediates as well as the synthetic processes described
herein.
[0038] In another aspect, the present invention provides synthetic
schemes and methods of use of compounds having Formula IA and
analogs or derivatives thereof. In another aspect, the invention
provides synthetic and modification schemes for preparing analogs
and derivatives of the compounds of Formula IA, as well as
compositions and methods for the use of such analogs and
derivatives.
[0039] The above summary of the present invention is not intended
to describe each disclosed embodiment or every implementation of
the present invention. The description that follows more
particularly exemplifies illustrative embodiments. In several
places throughout the application, guidance is provided through
lists of examples, which examples can be used in various
combinations. In each instance, the recited list serves only as a
representative group and should not be interpreted as an exclusive
list.
[0040] The details of one or more embodiments of the invention are
set forth in the accompanying description below. Other features,
objects, and advantages of the invention will be apparent from the
description and drawings, and from the claims.
BRIEF DESCRIPTION OF THE DRAWINGS
[0041] FIGS. 1A-1P illustrate syntheses of compounds of Formula
IA.
[0042] FIGS. 2A and 2B illustrate the effect of various
concentrations of a compound of Formula 1A on S1P (2A) and
sphingosine (Sph) and dihydroSph (2B) accumulation in U937 cells;
ca. 5 million cells per point, treatment time 2 hours.
[0043] FIG. 2C illustrates accumulation of a test compound of
Formula IA in U937 cells (about 100.times. concentration from
surrounding media) after 2 hours of treatment.
[0044] FIGS. 3A and 3B illustrate the S1P levels after 24 hours of
application of a test compound of Formula IA, e.g., very low levels
of sphinganine 1-phosphate (dhS1P, 3B) in U937 cells--off scale on
3A.
[0045] FIGS. 4A and 4B illustrate treated U937 cells at times, as
soon as possible (asap), 2 hours, or 2 hours/wash, then incubate 24
hours.
[0046] FIG. 4C illustrates retention of a compound of the Formula
IA in U937 cells after 24 hours in drug free media.
[0047] FIG. 5A illustrates the shows effect on synthesis of S1P by
a test compound of the Formula IA, in this instance VPC96091. Cells
were supplemented with exogenous sphingosine (Sph) at noted
concentrations and intracellular S1P was measured with, or without
addition of the test compound.
[0048] FIG. 5B illustrates the converse effect of that illustrated
in slide 5A, i.e., accumulation of intracellular Sph in cells
treated with Sph and a test compound of the Formula IA, in this
instance VPC96091.
[0049] FIG. 6A illustrates the persistence of test compound
VPC96091 in whole blood after IV injection of 10 mg/kg (mpk) in
rats.
[0050] FIGS. 6B and 6C illustrate the change (decrease) in levels
of enzyme products, S1P (6B) or dhS1P (6C), in whole rat blood
following IV injection of 10 mpk of test compound VPC96091.
[0051] FIGS. 6D and 6E illustrate the inverse change (increase) in
levels of enzyme substrates, Sph (6D) or dhSph (6E), in whole rat
blood following IV injection of 10 mpk VPC96091.
[0052] FIGS. 7A-7D illustrate the effect of dosing the amidoxime
pro-drug, VPC96143, in three mice (#51, 52, 53) at 20 mpk. FIG. 7A
illustrates plasma S1P levels. FIG. 7B illustrates accumulation in
plasma of the amidine congener, VPC96077, of the amidoxime,
VPC96143, and thus the conversion of the amidoxime to the amidine
in vivo. FIG. 7C illustrates tissue (liver, kidney) S1P levels.
FIG. 7D illustrates the ratio of amidine:amidoxime in liver and
kidney.
[0053] FIGS. 8A, 8B, 9A, and 9B illustrate the effect on S1P levels
and pro-drug conversion using human monocyte U937 cells.
DETAILED DESCRIPTION
[0054] The following abbreviations are used herein: sphingosine
kinase (types 1 and 2): SphK, sphingosine 1-phosphate, S1P, and
sphinganine 1-phosphate (dhS1P).
[0055] In describing and claiming the invention, unless otherwise
defined, all technical and scientific terms used herein have the
same meaning as commonly understood by one of ordinary skill in the
art to which this invention belongs. Although any materials and
methods similar or equivalent to those described herein can be used
in the practice or testing of the present invention, the preferred
materials and methods are described herein. Each of the following
terms has meaning associated with it in this section. Exemplary and
preferred values listed below for radicals, substituents, and
ranges are for illustrations only; they do not exclude other
defined values or other values within defined ranges for the
radicals and substituents.
[0056] The terms "a," "an," "the," "at least one," and "one or
more" are used interchangeably. Thus, for example, a composition
that comprises "an" element means one element or more than one
element.
[0057] The term "receptor agonists" are compounds that mimic the
action of S1P at one or more of its receptors but may have
differing potency and/or efficacy.
[0058] The term "receptor antagonists" are compounds that 1) lack
intrinsic agonist activity and 2) block agonist (e.g., S1P)
activation of the S1P receptor(s), often in a manner that is both
fully surmountable and reversible (`competitive antagonist`).
[0059] The term "affected cell" refers to a cell of a subject
afflicted with a disease or disorder, which affected cell has an
altered phenotype relative to a subject not afflicted with a
disease or disorder.
[0060] Cells or tissue are "affected" by a disease or disorder if
the cells or tissue have an altered phenotype relative to the same
cells or tissue in a subject not afflicted with a disease or
disorder.
[0061] A disease or disorder is "alleviated" if the severity of a
symptom of the disease or disorder, the frequency with which such a
symptom is experienced by a patient, or both, is reduced.
[0062] An "analog" of a chemical compound is a compound that, by
way of example, resembles another in structure but is not
necessarily an isomer (e.g., 5-fluorouracil is an analog of
thymine).
[0063] The terms "cell," "cell line," and "cell culture" may be
used interchangeably.
[0064] A "control" cell, tissue, sample, or subject is a cell,
tissue, sample, or subject of the same type as a test cell, tissue,
sample, or subject. The control may, for example, be examined at
precisely or nearly the same time the test cell, tissue, sample, or
subject is examined. The control may also, for example, be examined
at a time distant from the time at which the test cell, tissue,
sample, or subject is examined, and the results of the examination
of the control may be recorded so that the recorded results may be
compared with results obtained by examination of a test cell,
tissue, sample, or subject. The control may also be obtained from
another source or similar source other than the test group or a
test subject, where the test sample is obtained from a subject
suspected of having a disease or disorder for which the test is
being performed.
[0065] A "test" cell, tissue, sample, or subject is one being
examined or treated.
[0066] A "pathoindicative" cell, tissue, or sample is one which,
when present, is an indication that the animal in which the cell,
tissue, or sample is located (or from which the tissue is obtained)
is afflicted with a disease or disorder. By way of example, the
presence of one or more breast cells in a lung tissue of an animal
is an indication that the animal is afflicted with metastatic
breast cancer.
[0067] A tissue "normally comprises" a cell if one or more of the
cell are present in the tissue in an animal not afflicted with a
disease or disorder.
[0068] The use of the word "detect" and its grammatical variants is
meant to refer to measurement of the species without
quantification, whereas use of the word "determine" or "measure"
with their grammatical variants are meant to refer to measurement
of the species with quantification. The terms "detect" and
"identify" are used interchangeably herein.
[0069] A "detectable marker" or a "reporter molecule" is an atom or
a molecule that permits the specific detection of a compound having
the marker in the presence of similar compounds without a marker.
Detectable markers or reporter molecules include, e.g., radioactive
isotopes, antigenic determinants, enzymes, nucleic acids available
for hybridization, chromophores, fluorophores, chemiluminescent
molecules, electrochemically detectable molecules, and molecules
that provide for altered fluorescence-polarization or altered
light-scattering.
[0070] A "disease" is a state of health of an animal wherein the
animal cannot maintain homeostasis, and wherein if the disease is
not ameliorated then the animal's health continues to
deteriorate.
[0071] A "disorder" in an animal is a state of health in which the
animal is able to maintain homeostasis, but in which the animal's
state of health is less favorable than it would be in the absence
of the disorder. Left untreated, a disorder does not necessarily
cause a further decrease in the animal's state of health.
[0072] An "effective amount" means an amount sufficient to produce
a selected effect. For example, an effective amount of an S1P
receptor antagonist is an amount that decreases the cell signaling
activity of the S1P receptor.
[0073] A "functional" molecule is a molecule in a form in which it
exhibits a property by which it is characterized. By way of
example, a functional enzyme is one which exhibits the
characteristic catalytic activity by which the enzyme is
characterized.
[0074] The term "inhibit" refers to the ability of a disclosed
compound to reduce or impede a described function. Inhibition is by
at least 10%, preferably by at least 25%, more preferably by at
least 50%, even more preferably by at least 75%, and most
preferably, the function is inhibited by at least 95%.
[0075] The term "selective" refers to the ability of the disclosed
compounds to inhibit one of the sphingosine kinase 1 or sphingosine
kinase 2 (SphK1 & SphK2) enzymes and not the other enzyme.
Preferably, the selective compound will have a Ki value for one
enzyme that is less than, by at least an order of magnitude (e.g.,
a ten-fold difference), the Ki value for the other enzyme or in
inhibition of one of the SphK enzymes over the other enzyme.
[0076] An "effective amount" means an amount sufficient to produce
a selected effect. For example, an effective amount of a
sphingosine kinase 1 or sphingosine kinase 2 (SphK1 & SphK2)
inhibitor is an amount that inhibits substrate (sphingosine)
binding of the sphingosine kinases and thereby the conversion of
sphingosine to S1P.
[0077] A "functional" molecule is a molecule in a form in which it
exhibits a property by which it is characterized. By way of
example, a functional enzyme is one that exhibits the
characteristic catalytic activity by which the enzyme is
characterized.
[0078] The term "inhibit" refers to the ability of a disclosed
compound to reduce or impede a described function. Inhibition is by
at least 10%, preferably by at least 25%, more preferably by at
least 50%, even more preferably by at least 75%, and most
preferably, the function is inhibited by at least 95%.
[0079] The term "instructional material" includes a publication, a
recording, a diagram, or any other medium of expression that can be
used to communicate the usefulness of the disclosed compounds in
the kit for effecting alleviation of the various diseases or
disorders recited herein. Optionally, or alternately, the
instructional material may describe one or more methods of
alleviating the diseases or disorders in a cell or a tissue of a
mammal. The instructional material of the kit may, for example, be
affixed to a container which contains a disclosed compound or be
shipped together with a container which contains the identified
compound. Alternatively, the instructional material may be shipped
separately from the container with the intention that the
instructional material and the compound be used cooperatively by
the recipient.
[0080] The term "parenteral" means not through the alimentary canal
but by some other route such as subcutaneous, intramuscular,
intraspinal, or intravenous.
[0081] The term "purified" and similar terms relate to the
isolation of a molecule or compound in a form that is substantially
free (at least 75% free, preferably 90% free, and most preferably
at least 95% free) from other components normally associated with
the molecule or compound in a native environment. The term
"purified" does not necessarily indicate that complete purity of
the particular molecules achieved during the process. A "very pure"
compound refers to a compound that is greater than 90% pure. A
"highly purified" compound refers to a compound that is greater
than 95% pure.
[0082] A "sample" refers preferably to a biological sample from a
subject, including, but not limited to, normal tissue samples,
diseased tissue samples, biopsies, blood, saliva, feces, semen,
tears, and urine. A sample can also be any other source of material
obtained from a subject, which contains cells, tissues, or fluid of
interest. A sample can also be obtained from cell or tissue
culture.
[0083] The term "standard," refers to something used for
comparison. For example, a standard can be a known standard agent
or compound which is administered or added to a control sample and
used for comparing results when measuring said compound in a test
sample. Standard can also refer to an "internal standard", such as
an agent or compound which is added at known amounts to a sample
and is useful in determining such things as purification or
recovery rates when a sample is processed or subjected to
purification or extraction procedures before a marker of interest
is measured.
[0084] A "subject" of analysis, diagnosis, or treatment is an
animal. Such animals include mammals, preferably a human.
[0085] A "therapeutic" treatment is a treatment administered to a
subject who exhibits signs of pathology for the purpose of
diminishing or eliminating those signs.
[0086] A "therapeutically effective amount" of a compound is that
amount of compound which is sufficient to provide a beneficial
effect to the subject to which the compound is administered.
[0087] The term "treating" includes prophylaxis of the specific
disorder or condition, or alleviation of the symptoms associated
with a specific disorder or condition or preventing or eliminating
said symptoms.
[0088] The disclosed compounds are generally named according to the
IUPAC or CAS nomenclature system. Abbreviations which are well
known to one of ordinary skill in the art may be used (e.g., "Ph"
for phenyl, "Me" for methyl, "Et" for ethyl, "h" for hour or hours,
"r.t." for room temperature, "THF" for tetrahydrofuran, and "rac"
for racemic mixture).
[0089] The values listed below for radicals, substituents, and
ranges, are for illustration only; they do not exclude other
defined values or other values within defined ranges for the
radicals and substituents. The disclosed compounds include
compounds of Formula IA having any combination of the exemplary
values, preferred values, and more preferred values described
herein.
[0090] The term "halogen" or "halo" includes bromo, chloro, fluoro,
and iodo. The term "haloalkyl", refers to an alkyl radical bearing
at least one halogen substituent, non-limiting examples include,
but are not limited to, chloromethyl, fluoroethyl or
trifluoromethyl and the like. The term "C.sub.1-C.sub.20 alkyl"
refers to a branched or linear alkyl group having from one to
twenty carbons. Non-limiting examples include, but are not limited
to, methyl, ethyl, n-propyl, iso-propyl, butyl, iso-butyl,
sec-butyl, tert-butyl, pentyl, hexyl, heptyl, octyl and the like.
The term "C.sub.2-C.sub.20 alkenyl", refers to an olefinically
unsaturated branched or linear group having from two to twenty
carbon atoms and at least one double bond. Typically,
C.sub.2-C.sub.20 alkenyl groups include, but are not limited to,
1-propenyl, 2-propenyl, 1,3-butadienyl, 1-butenyl, hexenyl,
pentenyl, hexenyl, heptenyl, octenyl and the like. The term
(C.sub.2-C.sub.20)alkynyl can be ethynyl, 1-propynyl, 2-propynyl,
1-butynyl, 2-butynyl, 3-butynyl, 1-pentynyl, 2-pentynyl,
3-pentynyl, 4-pentynyl, 1-hexynyl, 2-hexynyl, 3-hexynyl, 4-hexynyl,
or 5-hexynyl, and the like. The term "(C.sub.1-C.sub.20)alkoxy"
refers to an alkyl group attached through an oxygen atom. Examples
of (C.sub.1-C.sub.20)alkoxy can be methoxy, ethoxy, propoxy,
isopropoxy, butoxy, iso-butoxy, sec-butoxy, pentoxy, 3-pentoxy,
hexyloxy, heptoxy, or octoxy, and the like. The term
(C.sub.2-C.sub.26)alkoxyalkyl can be methoxy methyl, methoxy ethyl,
ethoxy methyl, ethoxy ethyl, and the like.
[0091] The term "C.sub.3-C.sub.12 cycloalkyl" includes groups
having one ring or multiple rings. Non-limiting examples include be
cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl,
cyclooctyl, adamantyl and the like.
[0092] The term "optionally substituted" refers to zero, one, two,
three, four, five, six or seven substituents, wherein the
substituents are each independently selected. Each of the
independently selected substituents may be the same or different
than other substituents. Non-limiting examples of the substituents
include halo, halo(C.sub.1-C.sub.10)alkyl, cyano, amino
(--NR.sup.aR.sup.b), (C.sub.1-C.sub.20)alkoxy,
(C.sub.2-C.sub.26)alkoxyalkyl, (C.sub.3-C.sub.12)cycloalkyl,
(C.sub.6-C.sub.10)aryl, (C.sub.7-C.sub.30)alkylaryl,
(C.sub.2-C.sub.10)heterocyclic, or (C.sub.4-C.sub.10)-heteroaryl;
wherein one or more of the carbon atoms in the Z alkyl groups can
be independently replaced with non-peroxide oxygen, sulfur or
NR.sup.c; where each R.sup.a, R.sup.b, or R.sup.c is independently
hydrogen or (C.sub.1-C.sub.7)alkyl.
[0093] The term "(C.sub.6-C.sub.14)aryl" refers to a mono or
bicyclic carbocyclic ring system having one or two aromatic rings
including, but not limited to, phenyl, benzyl, naphthyl,
tetrahydronaphthyl, indanyl, indenyl, and the like.
[0094] The term "optionally substituted aryl" includes aryl
compounds having zero, one, two, three or four substituents, and a
substituted aryl includes aryl compounds having one, two, three or
four substituents, wherein the substituents include groups such as,
for example, alkyl, halo, amino or those substituents listed
herein.
[0095] The term "(C.sub.6-C.sub.14)aryl(C.sub.1-C.sub.20)alkyl",
"(C.sub.7-C.sub.30)arylalkyl" or "aralkyl" refers to an alkyl group
substituted with a mono or bicyclic carbocyclic ring system having
one or two aromatic rings including, a group such as phenyl,
naphthyl, tetrahydronaphthyl, indanyl, indenyl, and the like.
Non-limiting examples of arylalkyl include benzyl, phenyl ethyl,
and the like.
[0096] The term "(C.sub.2-C.sub.10)heterocyclic group" refers to an
optionally substituted mono- or bicyclic carbocyclic ring system
containing one, two, or three heteroatoms (optionally in each ring)
wherein the heteroatoms are non-peroxide oxygen, sulfur, and
nitrogen (NR).
[0097] The term "(C.sub.4-C.sub.10)heteroaryl" refers to an
optionally substituted mono- or bicyclic carbocyclic ring system
containing one, two, or three heteroatoms (optionally in each ring)
wherein the heteroatoms are non-peroxide oxygen, sulfur, and
nitrogen (NR.sup.c). Non-limiting examples of heteroaryl groups
include furyl, thienyl, pyridyl, and the like.
[0098] The term "bicyclic" represents either an unsaturated or
saturated stable bridged or fused bicyclic carbon ring. The
bicyclic ring may be attached at any carbon atom which affords a
stable structure. Typically a bicyclic ring system can have from
about 7 to about 12 atoms in the ring system. The term includes,
but is not limited to, naphthyl, dicyclohexyl, dicyclohexenyl, and
the like.
[0099] A "derivative" of a compound refers to a chemical compound
that may be produced from another compound of similar structure in
one or more steps, such as replacement of hydrogen by an alkyl,
acyl, or amino group.
[0100] The term "pharmaceutically acceptable carrier" includes any
of the standard pharmaceutical carriers, such as a phosphate
buffered saline solution, hydroxypropyl beta-cyclodextrins
(HO-propyl beta cyclodextrins), water, emulsions such as an
oil/water or water/oil emulsion, and various types of wetting
agents. The term also encompasses any of the agents approved by a
regulatory agency of the U.S. Federal government or listed in the
US Pharmacopeia for use in animals, including humans.
[0101] The term "pharmaceutically-acceptable salt" refers to salts
which retain the biological effectiveness and properties of the
disclosed compounds and which are not biologically or otherwise
undesirable. In many cases, the disclosed compounds are capable of
forming acid or base salts by virtue of the presence of amino or
carboxyl groups or groups similar thereto.
[0102] An "effective amount" means an amount sufficient to produce
a selected effect. For example, an effective amount of an S1P
receptor agonist is an amount that decreases the cell signaling
activity of the S1P receptor.
[0103] The disclosed compounds can contain one or more asymmetric
centers in the molecule. In accordance with the present disclosure
any structure that does not designate the stereochemistry is to be
understood as embracing all the various optical isomers, as well as
racemic mixtures thereof.
[0104] The disclosed compounds may exist in tautomeric forms and
the invention includes both mixtures and separate individual
tautomers. For example, the following structure:
##STR00002##
is understood to represent a mixture of the structures:
##STR00003##
as well as
##STR00004##
[0105] The terms 16:0, 18:0, 18:1, 20:4 or 22:6 hydrocarbon refers
to a branched or straight alkyl or alkenyl group, wherein the first
integer represents the total number of carbons in the group and the
second integer represent the number of double bonds in the
group.
[0106] The disclosed compounds can contain one or more asymmetric
centers in the molecule. In accordance with the present disclosure
any structure that does not designate the stereochemistry is to be
understood as embracing all the various optical isomers, as well as
racemic mixtures thereof.
[0107] It will be appreciated by those skilled in the art that the
disclosed compounds having chiral centers may exist in and be
isolated in optically active and racemic forms. It is to be
understood that the disclosed compounds encompass any racemic,
optically active or stereoisomeric form, or mixtures thereof, of
the compound, which possess the useful properties described herein,
such as the S,R; S,S; R,R; or R,S diastereomers. It is well known
in the art how to prepare such optically active forms (for example,
resolution of the racemic form by recrystallization techniques,
synthesis from optically-active starting materials, chiral
synthesis, or by chromatographic separation using a chiral
stationary phase) and how to determine sphingosine kinase activity
using the standard tests described herein, or using other similar
tests which are well known in the art. In addition, some compounds
may exhibit polymorphism.
[0108] Potential uses of SphK inhibitors include, but are not
limited to, anti-angiogenesis, treating neoplastic disease,
preventing restenosis, treating autoimmune disorders and treating
vascular injury.
[0109] The compounds of Formula IA are useful for treating a
disease or disorder including administering to a subject in need
thereof of a therapeutically acceptable amount of a compound of
Formula IA, or a pharmaceutical composition comprising a
therapeutically effective amount of a compound of Formula IA, and a
pharmaceutically-acceptable carrier.
[0110] The disclosed compounds are directed to improved SphK
inhibiting compounds and derivatives thereof, where, for example,
the compounds have been modified to enhance their oral availability
and thus increase their efficacy in oral pharmaceutical
formulations. The compounds of the present invention are believed
to act as inhibitors of SphK enzymes. More particularly, the
pro-drug derivatives, amidoximes of the present invention hydroxy
derivatives that have enhanced oral availability relative to the
parent compound. As shown herein the amidoximes (administered
orally) can be reduced to amidines by cells and tissues.
[0111] Exemplary and preferred values listed below for radicals,
substituents, and ranges, are for illustration only; they do not
exclude other defined values or other values within defined ranges
for the radicals and substituents.
[0112] Exemplary values for R.sup.1, hydrogen, or hydroxy.
[0113] Exemplary values for R.sup.2 and R.sup.3 are independently
hydrogen, methyl, or ethyl.
[0114] Additional exemplary values for R.sup.2 and R.sup.3 are
hydrogen.
[0115] Exemplary values for X and Y are independently NH and
C.dbd.O.
[0116] Additional exemplary values for X and Y together are
--C(.dbd.O)NR.sup.8-- or --NR.sup.c(.dbd.O)--.
[0117] Exemplary values for R include hydrogen, methyl, ethyl,
propyl or isopropyl.
[0118] Additional exemplary values for R.sup.4 and R.sup.5 together
with the atom to which they are attached include cyclopropyl,
cyclobutyl, cyclopentyl, or cyclohexyl.
[0119] Additional exemplary values for R.sup.4 and R.sup.5 together
with the atom to which they are attached include cyclopropyl.
[0120] Exemplary values for R.sup.4 and R.sup.8 together with the
atom to which they are attached include cyclopentyl, cyclohexyl or
pyrrolidinyl.
[0121] Additional exemplary values for R.sup.4 and R.sup.8 together
with the atom to which they are attached include pyrrolidinyl.
[0122] Exemplary values for Z include (C.sub.10-C.sub.18)alkyl,
(C.sub.10-C.sub.18)alkenyl, or (C.sub.7-C.sub.18)alkylphenyl.
[0123] Additional exemplary values for Z include compounds where
the alkyl chain in the alkylphenyl group has 9, 10, 11, 12, 13, 14,
15, or 16, carbon atoms.
[0124] Additional exemplary values for Z include
(C.sub.5-C.sub.17)alkylphenyl,
##STR00005## ##STR00006##
[0125] Exemplary heteroatom groups for replacing carbon atoms in a
ring or alkyl include O or NR.sup.10.
[0126] Exemplary values for R.sup.10 include hydrogen, methyl,
ethyl, propyl or isopropyl.
[0127] Exemplary compounds of the invention have formulas shown in
table 1 or salts thereof (the compounds are shown as amidoximes or
hydrochloride salts of the claimed compounds). Table 1 includes:
Inhibitory constants (Ki) of test compounds, all values in
micromolar concentrations. ND not determined
TABLE-US-00001 TABLE 1 Amidine Amidine No. Amidoxime (HCl salt)
SphK1 (.mu.M) SphK2 (.mu.M) VPC96031 ##STR00007## ##STR00008## 25
23 VPC95287 ##STR00009## ##STR00010## 5 4 VPC96047 ##STR00011##
##STR00012## 0.2 0.5 VPC96021 ##STR00013## ##STR00014## 10 32
VPC96055 ##STR00015## ##STR00016## 36 10 VPC171117 ##STR00017##
##STR00018## >100 90 VPC171147 ##STR00019## ##STR00020## 60 20
VPC171159 ##STR00021## ##STR00022## NA NA VPC171167 ##STR00023##
##STR00024## 4 10 VPC171137 ##STR00025## ##STR00026## 40 45
VPC143033 ##STR00027## ##STR00028## 3 4 VPC143081 ##STR00029##
##STR00030## 0.4 5 VPC143064 ##STR00031## ##STR00032## 0.3 6
VPC143065 ##STR00033## ##STR00034## 8.4 >100 VPC143066
##STR00035## ##STR00036## 30 >100 VPC143078 ##STR00037##
##STR00038## 5 38 VPC143129 ##STR00039## ##STR00040## 0.5 >10
VPC143119 ##STR00041## ##STR00042## 1.3 >10 VPC143113
##STR00043## ##STR00044## 0.39 12 VPC143105 ##STR00045##
##STR00046## 0.24 7 VPC143126 ##STR00047## ##STR00048## 0.8 >20
VPC143144 ##STR00049## ##STR00050## 0.11 26 VPC14a1058 ##STR00051##
##STR00052## 0.45 25 VPC143154 ##STR00053## ##STR00054## 0.17 18
VPC14a1051 ##STR00055## ##STR00056## 0.13 15 VPC143090 ##STR00057##
##STR00058## 1.5 4.6 VPC143057 ##STR00059## ##STR00060## 6 7.5
VPC143046 ##STR00061## ##STR00062## 5.4 >100 VPC14a1002
##STR00063## ##STR00064## 9 >100 VPC95291 ##STR00065##
##STR00066## 0.75 2.6 VPC95301 ##STR00067## ##STR00068## 8 50
VPC95127 ##STR00069## ##STR00070## >100 60 VPC95297 ##STR00071##
##STR00072## 5 4 VPC96077 ##STR00073## ##STR00074## 0.3 2 VPC96091
##STR00075## ##STR00076## 0.1 1.5 VPC96115 ##STR00077##
##STR00078## 16 5 VPC143237 ##STR00079## ##STR00080## 0.075 15
VPC143v47 ##STR00081## ##STR00082## NA NA VPC143v48 ##STR00083##
##STR00084## NA NA
[0128] Pharmaceutically acceptable base addition salts can be
prepared from inorganic and organic bases. Salts from inorganic
bases, include but are not limited to, sodium, potassium, lithium,
ammonium, calcium and magnesium salts. Salts derived from organic
bases include, but are not limited to, salts of primary, secondary
and tertiary amines, such as alkyl amines, dialkyl amines, trialkyl
amines, substituted alkyl amines, di(substituted alkyl) amines,
tri(substituted alkyl) amines, alkenyl amines, dialkenyl amines,
trialkenyl amines, substituted alkenyl amines, di(substituted
alkenyl) amines, tri(substituted alkenyl) amines, cycloalkyl
amines, di(cycloalkyl) amines, tri(cycloalkyl) amines, substituted
cycloalkyl amines, disubstituted cycloalkyl amine, trisubstituted
cycloalkyl amines, cycloalkenyl amines, di(cycloalkenyl) amines,
tri(cycloalkenyl) amines, substituted cycloalkenyl amines,
disubstituted cycloalkenyl amine, trisubstituted cycloalkenyl
amines, aryl amines, diaryl amines, triaryl amines, heteroaryl
amines, diheteroaryl amines, triheteroaryl amines, heterocyclic
amines, diheterocyclic amines, triheterocyclic amines, mixed di-
and tri-amines where at least two of the substituents on the amine
are different and are alkyl, substituted alkyl, alkenyl,
substituted alkenyl, cycloalkyl, substituted cycloalkyl,
cycloalkenyl, substituted cycloalkenyl, aryl, heteroaryl, or
heterocyclic and the like. Also included are amines where the two
or three substituents, together with the amino nitrogen, form a
heterocyclic or heteroaryl group. Mon-limiting examples of amines
include, isopropylamine, trimethyl amine, diethyl amine,
tri(iso-propyl) amine, tri(n-propyl) amine, ethanolamine,
2-dimethylaminoethanol, tromethamine, lysine, arginine, histidine,
caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine,
glucosamine, N-alkylglucamines, theobromine, purines, piperazine,
piperidine, morpholine, N-ethylpiperidine, and the like. It should
also be understood that other carboxylic acid derivatives would be
useful, for example, carboxylic acid amides, including
carboxamides, lower alkyl carboxamides, dialkyl carboxamides, and
the like.
[0129] In cases where compounds of Formula IA are sufficiently
basic or acidic to form stable nontoxic acid or base salts,
preparation and administration of the compounds, as
pharmaceutically acceptable salts may be appropriate. Examples of
pharmaceutically acceptable salts are organic acid addition salts
formed with acids which form a physiological acceptable anion, for
example, tosylate, methanesulfonate, acetate, citrate, malonate,
tartarate, succinate, benzoate, ascorbate, .alpha.-ketoglutarate,
and .alpha.-glycerophosphate. Inorganic salts may also be formed,
including hydrochloride, sulfate, nitrate, bicarbonate, and
carbonate salts.
[0130] The compounds of the formulas above include all enantiomers
thereof.
[0131] Pharmaceutically acceptable salts may be obtained using
standard procedures well known in the art, for example by reacting
a sufficiently basic compound such as an amine with a suitable acid
affording a physiologically acceptable anion. Alkali metal (for
example, sodium, potassium or lithium) or alkaline earth metal (for
example calcium) salts of carboxylic acids can also be made.
[0132] The compounds of Formula IA can be formulated as
pharmaceutical compositions and administered to a mammalian host,
such as a human patient in a variety of forms adapted to the chosen
route of administration, e.g., orally or parenterally, by
intravenous, intramuscular, topical or subcutaneous routes.
[0133] Thus, the present compounds may be systemically
administered, e.g., orally, in combination with a pharmaceutically
acceptable vehicle such as an inert diluent or an assimilable
edible carrier. They may be enclosed in hard or soft shell gelatin
capsules, may be compressed into tablets, or may be incorporated
directly with the food of the patient's diet. For oral therapeutic
administration, the active compound may be combined with one or
more excipients and used in the form of ingestible tablets, buccal
tablets, troches, capsules, elixirs, suspensions, syrups, wafers,
and the like. Such compositions and preparations should contain at
least about 0.1% of active compound. The percentage of the
compositions and preparations may, of course, be varied and may
conveniently be between about 2 to about 60% of the weight of a
given unit dosage form. The amount of active compound in such
therapeutically useful compositions is such that an effective
dosage level will be obtained.
[0134] The tablets, troches, pills, capsules, and the like may also
contain the following: binders such as gum tragacanth, acacia, corn
starch or gelatin; excipients such as dicalcium phosphate; a
disintegrating agent such as corn starch, potato starch, alginic
acid and the like; a lubricant such as magnesium stearate; and a
sweetening agent such as sucrose, fructose, lactose or aspartame or
a flavoring agent such as peppermint, oil of wintergreen, or cherry
flavoring may be added. When the unit dosage form is a capsule, it
may contain, in addition to materials of the above type, a liquid
carrier, such as a vegetable oil or a polyethylene glycol. Various
other materials may be present as coatings or to otherwise modify
the physical form of the solid unit dosage form. For instance,
tablets, pills, or capsules may be coated with gelatin, wax,
shellac or sugar and the like. A syrup or elixir may contain the
active compound, sucrose or fructose as a sweetening agent, methyl
and propylparabens as preservatives, a dye and flavoring such as
cherry or orange flavor. Of course, any material used in preparing
any unit dosage form should be pharmaceutically acceptable and
substantially non-toxic in the amounts employed. In addition, the
active compound may be incorporated into sustained-release
preparations and devices.
[0135] The active compound may also be administered intravenously
or intraperitoneally by infusion or injection. Solutions of the
active compound or its salts can be prepared in water, optionally
mixed with a nontoxic surfactant. Dispersions can also be prepared
in glycerol, liquid polyethylene glycols, triacetin, and mixtures
thereof and in oils. Under ordinary conditions of storage and use,
these preparations contain a preservative to prevent the growth of
microorganisms.
[0136] Exemplary pharmaceutical dosage forms for injection or
infusion can include sterile aqueous solutions or dispersions or
sterile powders comprising the active ingredient which are adapted
for the extemporaneous preparation of sterile injectable or
infusible solutions or dispersions, optionally encapsulated in
liposomes. In all cases, the ultimate dosage form should be
sterile, fluid and stable under the conditions of manufacture and
storage. The liquid carrier or vehicle can be a solvent or liquid
dispersion medium comprising, for example, water, ethanol, a polyol
(for example, glycerol, propylene glycol, liquid polyethylene
glycols, and the like), vegetable oils, nontoxic glyceryl esters,
and mixtures thereof. The proper fluidity can be maintained, for
example, by the formation of liposomes, by the maintenance of the
required particle size in the case of dispersions or by the use of
surfactants. The prevention of the action of microorganisms can be
brought about by various antibacterial and antifungal agents, for
example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal,
and the like. In many cases, it will be preferable to include
isotonic agents, for example, sugars, buffers or sodium chloride.
Prolonged absorption of the injectable compositions can be brought
about by the use in the compositions of agents delaying absorption,
for example, aluminum monostearate and gelatin.
[0137] Sterile injectable solutions are prepared by incorporating
the active compound in the required amount in the appropriate
solvent with various of the other ingredients enumerated above, as
required, followed by filter sterilization. In the case of sterile
powders for the preparation of sterile injectable solutions, the
preferred methods of preparation are vacuum drying and the freeze
drying techniques, which yield a powder of the active ingredient
plus any additional desired ingredient present in the previously
sterile-filtered solutions.
[0138] For topical administration, the present compounds may be
applied in pure form, e.g., when they are liquids. However, it will
generally be desirable to administer them to the skin as
compositions or formulations, in combination with a
dermatologically acceptable carrier, which may be a solid or a
liquid.
[0139] Exemplary solid carriers include finely divided solids such
as talc, clay, microcrystalline cellulose, silica, alumina and the
like. Useful liquid carriers include water, alcohols or glycols or
water-alcohol/glycol blends, in which the present compounds can be
dissolved or dispersed at effective levels, optionally with the aid
of non-toxic surfactants. Adjuvants such as fragrances and
additional antimicrobial agents can be added to optimize the
properties for a given use. The resultant liquid compositions can
be applied from absorbent pads, used to impregnate bandages and
other dressings, or sprayed onto the affected area using pump-type
or aerosol sprayers.
[0140] Thickeners such as synthetic polymers, fatty acids, fatty
acid salts and esters, fatty alcohols, modified celluloses or
modified mineral materials can also be employed with liquid
carriers to form spreadable pastes, gels, ointments, soaps, and the
like, for application directly to the skin of the user.
[0141] Examples of useful dermatological compositions which can be
used to deliver the compounds of Formula IA to the skin are known
to the art; for example, see Jacquet et al. (U.S. Pat. No.
4,608,392), Geria (U.S. Pat. No. 4,992,478), Smith et al. (U.S.
Pat. No. 4,559,157) and Wortzman (U.S. Pat. No. 4,820,508).
[0142] Useful dosages of the compounds of Formula IA can be
determined by comparing their in vitro activity, and in vivo
activity in animal models. Methods for the extrapolation of
effective dosages in mice, and other animals, to humans are known
to the art; for example, see U.S. Pat. No. 4,938,949.
[0143] Generally, the concentration of the compound(s) of Formula
IA in a liquid composition, such as a lotion, will be from about
0.1 to about 25 weight percent, preferably from about 0.5-10 weight
percent. The concentration in a semi-solid or solid composition
such as a gel or a powder will be about 0.1-5 weight percent
preferably about 0.5-2.5 weight percent based on the total weight
of the composition.
[0144] The amount of the compound, or an active salt or derivative
thereof, required for use in treatment will vary not only with the
particular salt selected but also with the route of administration,
the nature of the condition being treated and the age and condition
of the patient and will be ultimately at the discretion of the
attendant physician or clinician.
[0145] In general, however, a suitable dose will be in the range of
from about 0.5 to about 100 mg/kg, e.g., from about 10 to about 75
mg/kg of body weight per day, such as 3 to about 50 mg per kilogram
body weight of the recipient per day, preferably in the range of 6
to 90 mg/kg/day, most preferably in the range of 15 to 60
mg/kg/day.
[0146] The compound is conveniently administered in unit dosage
form; for example, containing 5 to 1000 mg, conveniently 10 to 750
mg, most conveniently, 50 to 500 mg of active ingredient per unit
dosage form.
[0147] Ideally, the active ingredient should be administered to
achieve peak plasma concentrations of the active compound of from
about 0.5 to about 75 .mu.M, preferably, about 1 to 50 .mu.M, most
preferably, about 2 to about 30 .mu.M. This may be achieved, for
example, by the intravenous injection of a 0.05 to 5% solution of
the active ingredient, optionally in saline, or orally administered
as a bolus containing about 1-100 mg of the active ingredient.
Desirable blood levels may be maintained by continuous infusion to
provide about 0.01-5.0 mg/kg/hr or by intermittent infusions
containing about 0.4-15 mg/kg of the active ingredient(s).
[0148] The desired dose may conveniently be presented in a single
dose or as divided doses administered at appropriate intervals, for
example, as two, three, four, or more sub-doses per day. The
sub-dose itself may be further divided, e.g., into a number of
discrete loosely spaced administrations; such as multiple
inhalations from an insufflator or by application of a plurality of
drops into the eye.
[0149] The disclosed method includes a kit comprising an inhibitor
compound of Formula IA and instructional material that describes
administering the inhibitor compound or a composition comprising
the inhibitor compound to a cell or a subject. This should be
construed to include other embodiments of kits that are known to
those skilled in the art, such as a kit comprising a (preferably
sterile) solvent for dissolving or suspending the inhibitor
compound or composition prior to administering the compound or
composition to a cell or a subject. Preferably, the subject is a
human.
[0150] The disclosed compounds and methods, as described above or
as discussed in the Examples below, there can be employed
conventional chemical, cellular, histochemical, biochemical,
molecular biology, microbiology, and in vivo techniques, which are
known to those of skill in the art. Such techniques are explained
fully in the literature.
[0151] Without further description, it is believed that one of
ordinary skill in the art can, using the preceding description and
the following illustrative examples, make and utilize the disclosed
compounds.
[0152] In another aspect the invention provides compounds having
the formula IB:
##STR00085##
[0153] Wherein:
[0154] R.sup.1 is H or OH;
[0155] R.sup.2 and R.sup.3 are independently H,
(C.sub.1-C.sub.4)alky; or halo(C.sub.1-C.sub.3)alkyl;
[0156] R.sup.4 is absent, H, NH.sub.2 or
(C.sub.1-C.sub.4)alkyl;
[0157] R.sup.5 is absent, H or (C.sub.1-C.sub.4)alkyl;
or R.sup.4R.sup.5 together are CH.sub.2--CH.sub.2 (cyclopropyl),
CH.sub.2--CH.sub.2--CH.sub.2 (cyclobutyl),
CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2 (cyclopentyl) or
CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2 (cyclohexyl);
[0158] X and Y are independently CR.sup.6R.sup.7, NR.sup.8, O, S or
C.dbd.O (carbonyl);
[0159] R.sup.6 and R.sup.7 are independently H or
(C.sub.1-C.sub.7)alkyl
[0160] R.sup.8 is H or (C.sub.1-C.sub.7)alkyl
or R.sup.6 and R.sup.7 together, or R.sup.8 are (C.sub.2-C.sub.4)
forming ring structures as shown;
[0161] Z is (C.sub.10-C.sub.20)alkyl, (C.sub.10-C.sub.20)alkenyl,
(C.sub.10-C.sub.20)alkynyl, (C.sub.10-C.sub.20)alkoxy,
(C.sub.10-C.sub.26)alkoxyalkyl, (C.sub.10-C.sub.12)cycloalkyl,
(C.sub.6-C.sub.10)aryl, (C.sub.6-C.sub.20)alkylaryl,
(C.sub.6-C.sub.20)alkoxylaryl (C.sub.7-C.sub.30)arylalkyl,
(C.sub.2-C.sub.10)heterocyclic, (C.sub.4-C.sub.10)heteroaryl, or
(C.sub.4-C.sub.10)heteroaryl(C.sub.1-C.sub.20)alkyl.
[0162] The invention includes pharmaceutically acceptable salts or
esters of the compounds of Formula IB.
[0163] Processes for preparing compounds of Formula IA or for
preparing intermediates useful for preparing compounds of Formula
IA are provided as further embodiments. Intermediates useful for
preparing compounds of Formula IA are also provided as further
embodiments. The processes are provided as further embodiments and
are illustrated in the schemes herein wherein the meanings of the
generic radicals are as given above unless otherwise qualified.
[0164] Processes for preparing compounds of Formula IA or for
preparing intermediates useful for preparing compounds of Formula
IA are provided as further embodiments of the invention.
Intermediates useful for preparing compounds of Formula IA are also
provided as further embodiments of the invention. The compounds of
the invention can be prepared using starting materials and methods
known in the art.
[0165] The invention is now described with reference to the
following Examples and Embodiments. Without further description, it
is believed that one of ordinary skill in the art can, using the
preceding description and the following illustrative examples, make
and utilize the disclosed compounds. The following working examples
therefore, are provided for the purpose of illustration only and
specifically point out the preferred embodiments, and are not to be
construed as limiting in any way the remainder of the disclosure.
Therefore, the examples should be construed to encompass any and
all variations which become evident as a result of the teaching
provided herein.
Examples--Materials and Methods
[0166] The following abbreviations are used throughout specific and
general experimental procedures: acetonitrile (MeCN), acetic acid
(AcOH), chloroform (CHCl.sub.3), ethyl acetate (EtOAc),
iso-propanol (i-PrOH), methanol (MeOH), trifluoroacetic acid (TFA),
water (H.sub.2O), hydrochloric acid (HCl), sodium sulfate
(Na.sub.2SO.sub.4), sodium carbonate (Na.sub.2CO.sub.3), sodium
bicarbonate (NaHCO.sub.3), Magnesium Sulfate (MgSO.sub.4),
potassium carbonate (K.sub.2CO.sub.3), phosphorus pentoxide
(P.sub.2O.sub.5), lithium hydroxide (LiOH), aqueous (aq.), hour
(h), minute (min), room temperature (r.t.).
[0167] Except as indicated otherwise, reactions were monitored by
thin layer chromatography (TLC) using 0.25 mm Whatman precoated
silica gel plates. Flash chromatography was performed with the
indicated solvents and Dynamic Adsorbents silica gel (particle size
0.023-0.040 mm).
[0168] All non-aqueous reactions were carried out in oven or
flame-dried glassware under an argon or nitrogen atmosphere with
dry solvents and magnetic stirring, unless otherwise stated. The
argon and nitrogen were dried by passing through a tube of
Drierite. Anhydrous diethyl ether (Et.sub.2O), toluene,
dichloromethane (CH.sub.2Cl.sub.2), methanol (MeOH), and
tetrahydrofuran (THF) and N,N-dimethylformamide (DMF) were
purchased from Aldrich or VMR Chemicals and used as received. THF
was dried over activated molecular sieves (4 .ANG.) prior to use.
All other reagents were purchased from Acros chemicals and Aldrich
chemicals.
[0169] Except as indicated otherwise, reactions were monitored by
thin layer chromatography (TLC) using 0.25 mm Whatman precoated
silica gel plates. Flash chromatography was performed with the
indicated solvents and Dynamic Adsorbents silica gel (particle size
0.023-0.040 mm).
[0170] Proton (.sup.1H) and carbon (.sup.13C) NMR spectra were
recorded on a Varian UnityInova 500/51 or Varian UnityInova 300/54
at 300K unless otherwise noted. Chemical shifts are reported in ppm
(.delta.) values relative to the solvent as follows: CDCl.sub.3
(.delta. 7.24 for proton and .delta. 77.0 for carbon NMR),
DMSO-d.sub.6 (.delta. 2.50 for proton and .delta. 39.5 for carbon
NMR) CD.sub.3OD (.delta. 3.31 for proton and .delta. 47.6 for
carbon NMR).
[0171] High performance liquid chromatography--mass spectrometry
(LCMS) was carried out on a Waters 2695 separations module and a
Finnigan LCQ series mass spectrometer. All compounds were evaluated
for purity using a Thompson Instrument Company Advantage C.sub.18
column. Columns were outfitted with 5 .mu.M beads with a 60 .ANG.
pore size; columns were 250 mm in length and 4.6 mm in diameter.
Mobile phase A consisted of HPLC grade H.sub.2O and 0.01% TFA;
mobile phase B consisted of MeCN and 0.01% TFA. LCMS identification
and purity utilized a binary gradient starting with 90% A and 10% B
and linearly increasing to 100% B over the course of 10 minutes,
followed by and isocratic flow of 100% B for an additional 10
minutes. A flow rate of 1 mL/min. was maintained throughout the
HPLC method. The purity of all products was determined by
integration of the total ion count (TIC) spectra and integration of
the ultraviolet (UV) spectra at 214 nm; a Waters 486 Tunable
Absorbance Detector was used to collect all UV data. Retention
times are abbreviated as t.sub.R; mass to charge ratios are
abbreviated as m/z.
General Experimental Procedures
General Procedure A: Schotten-Baumen Protection of Free Amino
Acids
[0172] To a stirring solution of 10% sodium carbonate (weight to
volume) in water (25 mL) was added the free amino acid (9.3 mmol).
To this solution was added either di-tert-butyl dicarbonate or
N-(Benzyloxycarbonyloxy)succinamide (18.6 mmol) followed by
1,4-dioxanes (18.6 mL). This solution was allowed to stir for
approximately 15 hours at which point the reaction mixture was
extracted with three 15 mL potions of diethyl ether. The aqueous
layer was then acidified to a pH of 3 and immediately extracted
with four to five 25 mL portions of ethyl acetate. The organic
layers were then combined and washed with a 10 mL portion of brine,
dried with MgSO.sub.4, and evaporated to dryness. The crude
material was immediately taken on with no further purification.
General Procedure B: Conversion of a Carboxylic Acid to a Primary
Amide
[0173] To the stirring solution of the carboxylic acid (9.3 mmol)
in 20 mL of DCM was added triethylamine (3.76 g). After cooling the
solution to -25.degree. C., iso-butylchloroformate (2.54 g) was
added dropwise to the mixture. This mixture was stirred for
approximately 45 minutes and analyzed by TLC. Upon completion of
the reaction, 3.5 mL of a 28% solution of ammonium hydroxide in
water was added to the reaction mixture (still at -25.degree. C.)
and stirred for an additional 30 minutes. After this time the
mixture was warmed to r.t. and stirred for an additional 15 hours
at this temperature. When the reaction was deemed complete by TLC,
the reaction mixture was evaporated to an aqueous solution, diluted
with 150 mL of EtOAc and extracted with four 15 mL potions of
de-ionized water. The organic layer was washed once with brine,
dried over MgSO.sub.4 and evaporated to dryness. The crude mixture
was taken on without further purification.
General Procedure C: Conversion of a Primary Amide to a Nitrile
[0174] The primary amide (9.3 mmol) and triethylamine (1.9 g) was
taken up in 93 mL of anhydrous THF and cooled to 0.degree. C. Upon
cooling, trifluoroacetic anhydride (2.34 g) was added drop-wise to
the reaction mixture; after 10 minutes of stirring at this low
temperature the reaction mixture was allowed to warm to ambient
temperature and evaporated to dryness. The crude organic mixture
was taken up in 200 mL of EtOAc and extracted with four 15 mL
portions of 1N HCl followed by one 10 mL portion of brine. The
organic layer was then dried of MgSO.sub.4 and evaporated to an
oil. The crude product was purified by flash chromatography and
identified by NMR analysis.
General Procedure D: De-Protection of N-Boc Protected
Amino-Nitriles
[0175] To a stirring solution of the N-Boc protected amino-nitrile
(1.2 mmol) in anhydrous DCM (12 mL) was added trifluoroacetic acid,
drop-wise (12 mL) at room temperature. After 15 minutes the
reaction mixture was evaporated to dryness. Three 5 mL portions of
methanol were added to the crude oil and evaporated immediately;
three 5 mL portions of diethyl ether were then added and evaporated
in the same fashion to provide the trifluoroacetate salt as either
a yellowish solid or oil, depending on the substrate. This salt was
carried on immediately in the same reaction vessel.
General Procedure E: PyBOP Mediated Couplings of HCl or TFA Salts
to Acids
[0176] To a suspension of either the HCl or TFA salt of an amine
(0.43 mmols) in anhydrous DCM (9 mL) was added the acid (0.43
mmols) and PyBOP (0.223 g). Finally, diisopropylethylamine (0.222
g) was added and the reaction was allowed to stir for 15 hours. At
this time the reaction mixture was evaporated to dryness and
reconstituted in 100 mLs of EtOAc. The solution was extracted with
four 15 mL portions of 1N HCl followed by one portion of brine. The
organic layer was dried with MgSO.sub.4 and evaporated to dryness.
The crude organic material was purified by flash
chromatography.
General Procedure F: Amide Formation from an Amine and an Acid
Chloride
[0177] To a solution of a primary amine salt (1.68 mmol) in DCM (17
mLs) stirring at 0.degree. C. was added diisopropylethyl amine
(5.04 mmol) followed by an acid chloride (2.184 mmols). The
reaction mixture was allowed to warm to ambient temperature slowly
and stirred for an additional 15 hours. At this time the reaction
mixture was evaporated to dryness and reconstituted in ethyl
acetate (150 mLs). The solution was next extracted with three 15 mL
equivalents of 1N HCl followed by one 15 mL equivalent of brine.
The organic layer was then dried with magnesium sulfate, filtered
through a fritted funnel and dried to an oil. The crude mixture was
purified via flash chromatography.
General Procedure G: Conversion of Carboxylic Acids to Acid
Chlorides
[0178] An acid (0.28 mmols) and a catalytic amount of
dimethylformamide were dissolved in dichloromethane (2.8 mLs) and
the solution was cooled to 0.degree. C. To this solution was added
oxalyl chloride (0.84 mmols) and the reaction mixture was warmed to
room temperature. After stirring for 2 hours, the reaction mixture
was evaporated to dryness; the crude material was taken up in 3 one
mL portions of diethyl ether and evaporated. The oil was dried
under vacuum for an additional 30 minutes and carried on.
General Procedure H: Direct Conversion of Nitriles to Amidines
[0179] A nitrile (0.21 mmols) was taken up in 2.1 mL of anhydrous
methanol and 42 .mu.L of a solution of 0.5 M sodium methoxide in
methanol was immediately added. This mixture was allowed to stir
for 15 hours at which time 0.231 mmols of ammonium chloride was
added to the reaction. When the reaction appeared complete by TLC,
the product was purified in one of two ways. The reaction mixture
was evaporated to dryness and the crude material was taken up in
chloroform and filtered through a fine fritted funnel. The eluent
was collected and thoroughly dried under vacuum. Then the reaction
mixture was taken up in Et.sub.2O and filtered again through a fine
fritted funnel; the product was recovered as a white or off-white
solid.
General Procedure I: Pinnick Oxidation of Aldehydes to Carboxylic
Acids
[0180] An aryl aldehyde (3.05 mmol) was taken up in a 78 mL
solution of 1:1 THF and tert-butyl alcohol followed by 1.71 g of
2-methyl-2-butene. In a separate flask 0.83 g of sodium chlorite
and 1.26 g of sodium phosphate monobasic were dissolved in 13 mL of
H.sub.2O. Once fully dissolved, the aqueous solution was poured
into the stirring organic solution and stirred vigorously. Upon
completion by TLC, the reaction mixture was evaporated to dryness
and reconstituted in 100 mLs of EtOAc. The organic solution was
extracted with three 15 mL portions of 1N HCl followed by a single
10 mL portion of brine. The organic layer was finally dried over
magnesium sulfate and evaporated to dryness. No further
purification was necessary.
General Procedure J: Ether Synthesis from an Alkylbromide and an
Alcohol
[0181] To a solution of an alcohol (13.12 mmol) in DMF (40 mL) at
0.degree. C. was added 60% NaH in mineral oil (13.12 mmol) and let
stir at that temperature for 45 min. The alkyl bromide (6.56 mmol)
was then added and the reaction let to warm in the ice bath for 12
hours. The reaction was then cooled to 0.degree. C. and slowly
quenched with 1N HCl. The mixture was then extracted into three 200
mL portions ethyl acetate, dried with Na.sub.2SO.sub.4, and
concentrated to a yellow oil. The title product was recovered after
purification by flash chromatography.
General Procedure K: Ether Synthesis from an Alkyl Bromide and a
Phenol
[0182] A phenol (8.2 mmols), alkyl bromide (12.3 mmols) and cesium
carbonate (16.4 mmols) were dissolved in 14.4 mL of DMF. The
reaction mixture was then heated to 80.degree. C. until the
starting phenol appeared consumed via TLC analysis. After this
time, the reaction mixture was diluted with 200 mL of EtOAc and the
solution was washed with eight 10 mL portions of H.sub.2O and two
10 mL portions of brine. The organic layer was then dried over
MgSO.sub.4 and evaporated to dryness. The title product was
recovered in quantitative yield after purification by flash
chromatography.
General Procedure L: Organocuprate Substitution of an
Alkylbromide
[0183] To a solution of cuprous chloride (0.33 mmol) in Et.sub.2O
(7 mL) at -78.degree. C. was added a 2.0M solution BnMgBr in THF
(13.1 mmol) followed by an alkyl bromide (6.52 mmol) and let warm
to r.t. After reacting at r.t. for 20 hours, the reaction was
cooled to 0.degree. C. and quenched with 1 n HCl. The mixture was
then extracted into three 200 mL portions ethyl acetate, dried with
Na.sub.2SO.sub.4, and concentrated to a black oil. The title
product was recovered after purification by flash
chromatography.
General Procedure M: Hydroboration and Suzuki Coupling of an Olefin
and Arylbromide
[0184] An aryl aldehyde (3.05 mmol) was taken up in a 78 mL
solution of 1:1 THF and tert-butyl alcohol followed by 1.71 g of
2-methyl-2-butene. In a separate flask 0.83 g of sodium chlorite
and 1.26 g of sodium phosphate monobasic were dissolved in 13 mL of
H.sub.2O. Once fully dissolved, the aqueous solution was poured
into the stirring organic solution and stirred vigorously. Upon
completion by TLC, the reaction mixture was evaporated to dryness
and reconstituted in 100 mLs of EtOAc. The organic solution was
extracted with three 15 mL portions of 1N HCl followed by a single
10 mL portion of brine. The organic layer was finally dried over
magnesium sulfate and evaporated to dryness. No further
purification was necessary.
(Alternate Pathway for Hydroboration and Suzuki Coupling of an
Olefin and Arylbromide.)
[0185] To a solution of alkene (4.52 mmol) in THF (23 mL) at r.t.
was added a 0.5 M solution of 9-BBN (18 mL) and let stir for 12
hrs. 3 M K.sub.3PO.sub.4 (3 mL) was then added to the reaction and
let stir for 10 mins. The reaction was then diluted with DMF (23
mL) followed by the addition of the aryl bromide (4.97 mmol) then
PdCl.sub.2(dppf) (0.23 mmol) and let stir at r.t. for 6 hrs. The
reaction was then extracted into three 200 mL portions ethyl
acetate, dried with Na.sub.2SO.sub.4, and concentrated to a black
oil. The product was recovered after purification by flash
chromatography.
General Procedure N: Zinc Mediated Reduction of a Nitrobenzene to
an Aniline
[0186] To a solution of a nitrobenzene (2.00 mmol) in AcOH (10 mL)
at r.t. was added Zn dust by solid addition and let stir for 12
hrs. The reaction was then diluted with ethyl acetate and filtered
through a pad of Celite.TM.. The filtrate was evaporated to dryness
and the product was recovered after purification by flash
chromatography.
General Procedure O: Hydroboration/Oxidation of a Terminal Olefin
to an Alcohol
[0187] To a solution of an alkene (4.3 mmol) in dry THF (2.9 mL) at
0.degree. C. was added 9-BBN (0.5 M in THF, 9.4 mL, 4.7 mmol). The
reaction was immediately brought to room temperature and allowed to
react for 2 hours. Upon total consumption of starting alkene the
solution cooled down to -5.degree. C. and charged with EtOH (2.7
mL), 3 N NaOH (1.4 mL) and 30% H.sub.2O.sub.2 (1.5 mL). The
reaction was brought immediately to room temperature and allowed to
stir for 2 h. The reaction is then diluted with EtOAc and washed
with 1 N HCl. The organic layer was dried with Na.sub.2SO.sub.4,
filtered and concentrated to a crude oil. Crude material was
subjected to flash chromatography.
General Procedure P: Azide Formation from a Primary Alcohol
[0188] To a solution of an alcohol (876 mg, 3.2 mmol) in DCM (32
mL) at 0.degree. C. was added triethylamine (4.8 mmol) dropwise
followed by methanesulfonyl chloride (3.5 mmol) dropwise. Reaction
was immediately brought to room temperature and allowed to react
for 2 hours. Upon consumption of starting material by TLC reaction
is evaporated and reconstituted in EtOAc. Crude solution is
filtered through silica gel and eluent is concentrated. Crude
material is diluted in DMF (32 mL) and NaN.sub.3 is added (32
mmol). Reaction is heated to 100.degree. C. and allowed to react
overnight. The reaction was quenched by the slow addition of brine
and then diluted with EtOAc. The organic layer was washed with
brine, dried, with Na.sub.2SO.sub.4, filtered and concentrated to a
crude oil. Crude material was subjected to flash
chromatography.
General Procedure Q: Azide Reduction to an Amine
[0189] To a solution of an azide (1.2 mmol) in EtOAc (12 mL) at
room temperature was added palladium on carbon (0.1 mmol) and
reaction fitted with a hydrogen balloon (1 atm). Reaction was
allowed to stir for 24 hours. Reaction is diluted with EtOAc and
filtered through a plug of Celite and concentrated. Crude material
is diluted in DMF (12 mL) and chilled to 0.degree. C.
1-cyano-1-cyclopropanecarboxylic acid (266 mg, 0.86 mmol), PyBOP
(1.87 g, 3.6 mmol), and diisopropyethylamine (627 .mu.L, 3.6 mmol)
are added. The reaction is allowed to warm to room temperature
overnight. The reaction was quenched by the slow addition of brine
and then diluted with EtOAc. The organic layer was washed with
brine, dried, with Na.sub.2SO.sub.4, filtered and concentrated to a
crude oil. Crude material was subjected to flash
chromatography.
Synthesis of Specific Intermediates
(S)--N-(1-cyanoethyl)oleamide (1)
[0190] General procedures D was used to deprotect 1.17 mmols of
(S)-tert-butyl 1-cyanoethylcarbamate to the corresponding amine.
General procedure F was then use to couple to amine to 1.521 mmols
of oleoyl chloride. After standard purification techniques, 0.62
mmols (53%) of the title product was recovered. .sup.1H NMR (500
MHz, CDCl.sub.3) .delta. 6.08 (s, 1H), 5.40-5.28 (m, 2H), 5.00-4.90
(m, 1H), 4.12 (q, J=7.2, OH), 2.20 (dd, J=6.6, 8.5, 2H), 2.00 (dd,
J=6.3, 12.0, 4H), 1.68-1.58 (m, 2H), 1.54 (d, J=7.2, 3H), 1.38-1.20
(m, 24H), 0.87 (t, J=6.9, 3H). .sup.13C NMR (126 MHz, CDCl.sub.3)
.delta. 172.49, 130.02, 129.66, 119.35, 36.11, 35.63, 31.88, 29.74,
29.68, 29.50, 29.31, 29.20, 29.14, 29.08, 27.20, 27.14, 25.30,
22.67, 19.43, 14.11.
N-(1-cyanocyclopropyl)-4-decylbenzamide (3)
[0191] General procedure E was used to couple 1.68 mmols of
1-amino-cyclpropanecarbonitrile and 1.68 mmols of 4-decylbenzoic
acid. The reaction mixture was heated to 50.degree. C. for 48
hours; after this time standard purification techniques were used
to recover 0.84 mmols (50%) of the title product. .sup.1H NMR (500
MHz, CDCl.sub.3) .delta. 7.70 (dd, J=3.2, 8.1, 2H), 7.21 (d, J=8.0,
2H), 2.66-2.58 (m, 2H), 1.59 (d, J=5.6, 4H), 1.36 (q, J=5.8, 2H),
1.32-1.20 (m, 15H), 0.87 (t, J=6.9, 3H). .sup.13C NMR (126 MHz,
CDCl.sub.3) .delta. 168.16, 148.04, 129.96, 128.74, 127.32, 120.30,
35.86, 31.88, 31.15, 29.59, 29.45, 29.31, 29.23, 22.67, 20.88,
16.91, 14.12.
VPC95287
[0192] General procedure H was used to convert 0.12 mmols of 3 to
the title product. After standard purification techniques, 0.02
mmols (17%) of the title product was recovered and submitted for
biological evaluation. .sup.1H NMR (500 MHz, CD.sub.3OD) .delta.
7.81 (d, J=6.1, 2H), 7.30 (d, J=6.3, 2H), 2.67 (t, J=6.7, 2H), 1.75
(s, 2H), 1.63 (s, 2H), 1.56 (s, 2H), 1.29 (d, J=20.9, 14H), 0.89
(dd, J=4.5, 7.0, 3H). .sup.13C NMR (126 MHz, CD.sub.3OD) .delta.
172.51, 169.93, 147.78, 130.20, 128.34, 128.04, 127.48, 35.32,
32.59, 31.63, 30.97, 29.27, 28.81, 28.66, 22.30, 18.15, 12.98.
t.sub.R=10.34 min.; m/z=344.5.
N-(1-cyanocyclopropyl)-4-octylbenzamide (4-C8)
[0193] General procedure G was used to convert 0.53 mmols of
4-octylbenzoic acid to an acid chloride. After standard
purification techniques, the acyl-chloride was isolated in
quantitative yield and carried on immediately. General procedure F
was used to couple 0.41 mmols of 1-amino-1-cyclopropanecarbonitrile
to 0.53 mmols of the acid chloride. Standard purification
techniques were used to isolate 0.21 mmols (51%) of the title
product. .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 7.74-7.67 (m,
2H), 7.21 (d, J=8.4, 2H), 7.12 (s, 1H), 2.68-2.58 (m, 2H), 1.59
(dd, J=5.8, 8.3, 4H), 1.35 (dd, J=5.9, 8.3, 2H), 1.33-1.21 (m,
10H), 0.87 (t, J=7.0, 3H). .sup.13C NMR (126 MHz, CDCl.sub.3)
.delta. 168.06, 148.02, 129.96, 128.73, 127.27, 120.23, 35.85,
31.83, 31.13, 29.39, 29.21, 22.63, 20.88, 16.91, 14.08.
VPC96031
[0194] General procedure H was used to convert 0.17 mmols of 4-C8
to the corresponding amidine. After standard purification
techniques, 0.04 mmols (24%) of the title product were recovered.
.sup.1H NMR (500 MHz, CD.sub.3OD) .delta. 7.82 (d, J=8.3, 2H), 7.30
(d, J=8.2, 2H), 2.76-2.62 (m, 2H), 1.75 (dd, J=5.9, 8.5, 2H), 1.64
(dd, J=7.1, 14.3, 2H), 1.56 (dd, J=6.1, 8.4, 2H), 1.29 (dd, J=7.1,
15.0, 11H), 0.89 (t, J=7.0, 3H). .sup.13C NMR (126 MHz, CD.sub.3OD)
.delta. 172.52, 169.94, 147.79, 130.21, 128.19, 127.47, 35.32,
32.59, 31.58, 30.98, 29.11, 28.97, 28.82, 22.29, 18.16, 12.99.
LCMS: t.sub.R=8.5 minutes; m/z=316.
N-(1-cyanocyclopropyl)-4-dodecylbenzamide (4-C12)
[0195] General procedure G was used to convert 4-dodecylbenzoic
acid (0.52 mmols) to the corresponding acyl chloride. After
standard work up procedures, the intermediate was immediately
subjected to conditions described in general procedure F in the
presence of 1-amino-1-cyclopropanecarbonitrile hydrochloride (0.4
mmols). After flash chromatography, 0.15 mmols of the title product
was recovered. .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 7.68 (d,
J=7.1, 2H), 7.23 (d, J=7.5, 2H), 6.82 (s, 1H), 2.63 (t, J=7.7, 2H),
1.65-1.57 (m, 4H), 1.35 (t, J=7.0, 2H), 1.29-1.25 (m, 17H), 0.88
(t, J=6.5, 3H). .sup.13C NMR (126 MHz, CDCl.sub.3) .delta. 167.81,
148.06, 130.01, 128.75, 127.19, 120.07, 35.85, 31.89, 31.11, 29.61,
29.53, 29.42, 29.32, 29.20, 22.66, 20.88, 16.97, 14.08.
VPC96047.
[0196] General procedure H was used to convert 4-C12 (0.15 mmols)
to the corresponding amidine. Upon previously described
recrystallization procedures 0.06 mmols (40%) of the title product
was recovered. .sup.1H NMR (500 MHz, DMSO) .delta. 9.08 (s, 1H),
8.66 (bs, 4H), 7.81 (d, J=6.7, 2H), 7.27 (d, J=6.9, 2H), 2.60 (s,
2H), 1.65 (s, 2H), 1.54 (s, 2H), 1.37 (s, 2H), 1.21 (s, 19H), 0.83
(s, 3H). .sup.13C NMR (126 MHz, DMSO) .delta. 172.22, 167.99,
147.01, 131.21, 128.45, 128.32, 35.38, 33.09, 31.74, 31.23, 29.45,
29.27, 29.16, 28.98, 22.54, 18.42, 14.42. LCMS: t.sub.R=10.85
minutes; m/z=372.
N-(1-cyanocyclopropyl)-4-tetradecylbenzamide (4-C14)
[0197] General procedure G was used to convert 0.61 mmols of
4-tetradecylbenzoic acid to an acid chloride. After standard
purification techniques, the acyl-chloride was isolated in
quantitative yield and immediately. General procedure F was used to
couple 0.47 mmols of 1-amino-1-cyclopropanecarbonitrile. After
standard purification techniques, 0.28 mmols (60%) of the title
product was recovered. .sup.1H NMR (300 MHz, CDCl.sub.3) .delta.
7.68 (d, J=8.2, 2H), 7.23 (d, J=8.2, 2H), 6.85 (s, 1H), 2.63 (t,
J=9, 2H); 1.64-1.55 (m, 4H), 1.35 (dd, J=5.8, 8.5, 2H), 1.25 (s,
20H), 0.87 (t, J=6.6, 3H). .sup.13C NMR (75 MHz, CDCl.sub.3)
.delta. 168.05, 148.30, 130.19, 128.98, 127.43, 120.32, 36.09,
32.15, 31.38, 29.88, 29.68, 29.58, 29.45, 22.92, 21.10, 17.20,
14.36.
VPC96021
[0198] General procedure H was used to convert 0.26 mmols of 4-C14
to the title product. After standard purification techniques, 0.04
mmols (15%) of product was recovered and submitted for biological
evaluation. .sup.1H NMR (500 MHz, DMSO) .delta. 9.04 (s, 1H), 8.61
(bs, 4H), 7.81 (d, J=7.8, 2H), 7.27 (d, J=5.8, 2H), 2.60 (t, J=7.1,
2H), 1.65 (s, 2H), 1.54 (d, J=5.5, 2H), 1.37 (s, 2H), 1.24-1.21 (m,
21H), 0.83 (t, J=6.5, 3H). .sup.13C NMR (126 MHz, DMSO) .delta.
172.23, 168.02, 147.03, 131.24, 128.45, 128.31, 127.93, 35.38,
33.12, 31.72, 31.20, 29.44, 29.25, 29.13, 28.98, 22.52, 18.40,
14.39. LCMS: t.sub.R=12.3 minutes; m/z=400.
N-(1-cyanocycloproyl)-4-hexadecylbenzamide (4-C16)
[0199] General procedure G was used to convert hexadecanoic acid
(0.58 mmols) to the corresponding acyl chloride. After standard
work up techniques, the intermediated was coupled to
1-amino-1-cyclopropanecarbonitrile hydrochloride using the methods
described in general procedure F. Upon flash chromatography 0.41
mmols of the title product was recovered. .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta. 7.68 (d, J=8.2, 2H), 7.23 (d, J=8.1, 2H), 6.81
(s, 1H), 2.63 (t, J=7.7, 3H), 1.64-1.58 (m, 4H), 1.35 (q, J=4.9,
2H), 1.33-1.21 (m, 27H), 0.88 (t, J=6.9, 3H). .sup.13C NMR (126
MHz, CDCl.sub.3) .delta. 167.79, 148.06, 130.01, 128.75, 127.19,
120.07, 97.26, 35.84, 31.90, 31.11, 29.66, 29.63, 29.54, 29.43,
29.33, 29.20, 22.67, 20.88, 16.96, 14.08.
VPC96055
[0200] General procedure H was used to convert 0.18 mmols of 16-C16
to the corresponding amidine. However, instead of methanol, ethanol
was used to solubilize the starting material. After standard
recrystallization techniques, 0.04 mmols (22%) of the title product
was recovered as a white solid. .sup.1H NMR (500 MHz, DMSO) .delta.
9.06 (s, 1H), 8.65 (bs, 4H), 7.81 (d, J=7.5, 2H), 7.27 (d, J=7.5,
2H), 2.67-2.56 (m, 2H), 1.65 (s, 2H), 1.54 (d, J=4.0, 2H), 1.37 (s,
2H), 1.21 (bs, 25H), 0.84 (t, J=5.4, 3H). .sup.13C NMR (126 MHz,
DMSO) .delta. 172.16, 168.01, 147.02, 131.21, 128.45, 128.31,
35.38, 33.11, 31.74, 31.24, 29.46, 29.27, 29.15, 28.99, 22.54,
18.41, 14.42. LCMS: t.sub.R=12.4; m/z=428.
4-(heptyloxy)benzaldehyde (5-C7)
[0201] General procedure K was used to convert
4-hydroxybenzaldehyde (8.2 mmols) and 1-bromoheptane (12.3 mmols)
to 6.6 mmols (80%) of the title product was recovered. .sup.1H NMR
(300 MHz, CDCl.sub.3) .delta. 9.87 (s, 1H), 7.82 (d, J=8.8 Hz, 2H),
6.99 (d, J=8.7 Hz, 2H), 4.03 (t, J=6.5 Hz, 2H), 1.81 (quin., J=6
Hz, 1H), 1.24-1.53 (m, 8H), 0.89 (t, J=6.8 Hz, 3H); .sup.13C NMR
(75 MHz, CDCl.sub.3) .delta. 191.0, 164.4, 132.13, 129.8, 114.9,
68.6, 31.9, 29.18, 29.1, 26.1, 22.7, 14.2.
4-(heptyloxy)benzoic acid (6-C7)
[0202] General procedure I was used to convert 5-C7 (1.5 g, 6.6
mmol) to 6-C7 (1.6 g, 6.6 mmol, 100%) .sup.1H NMR (300 MHz,
DMSO-d.sub.6) .delta. 7.87 (d, J=8.7 Hz, 2H), 6.99 (d, J=8.8 Hz,
2H), 4.01 (t, J=6.4 Hz, 2H), 1.71 (quin., J=6.5 Hz, 2H), 1.47-1.19
(m, 4H), 0.86 (t, J=6.5 Hz, 3H); .sup.13C NMR (75 MHz,
DMSO-d.sub.6) .delta. 167.0, 162.3, 131.3, 122.8, 114.17, 67.8,
31.2, 28.5, 28.4, 25.4, 22.0, 13.9.
N-(1-cyanocyclopropyl)-4-(heptyloxy)benzamide (7-C7)
[0203] General procedure G was used to convert 1.00 mmols of 6-C7
to an acid chloride. After standard purification techniques, the
acyl-chloride was isolated in quantitative yield and immediately.
General procedure F was used to couple 0.84 mmols of
1-amino-1-cyclopropanecarbonitrile. After standard purification
techniques, 0.20 mmols (24%) of the title product was recovered.
.sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 7.77 (d, J=8.8 Hz, 2H),
6.85 (d, J=8.8 Hz, 2H), 3.94 (t, J=6.5 Hz, 2H), 1.76 (quin., J=5
Hz, 2H), 1.53 (t, J=5.7 Hz, 2H), 1.25-1.45 (m, 8H), 0.88 (t, J=6.7
Hz, 3H); .sup.13C NMR (75 MHz, CDCl.sub.3) .delta. 168.2, 162.6,
129.4, 124.6, 120.7, 114.4, 68.3, 31.8, 29.8, 29.2, 29.1, 26.0,
22.7, 21.0, 16.88, 14.2.
VPC171117
[0204] General procedure H was used to convert 0.20 mmols of 7-C7
to the title product. After standard purification techniques, 0.13
mmols (65%) of product was recovered and submitted for biological
evaluation. .sup.1H NMR (500 MHz, CD.sub.3OD) .delta.7.87 (d, J=8.6
Hz, 2H), 6.98 (d, J=8.7 Hz, 2H), 4.04 (t, J=6.4 Hz, 2H), 1.84-1.71
(m, 4H), 1.56 (bs, 2H), 1.52-1.43 (quin., J=7.5 Hz, 2H), 1.41-1.32
(m, 6H), 0.92 (t, J=6.8 Hz, 3H); .sup.13C NMR (125 MHz, CD.sub.3OD)
.delta. 174.0, 170.9, 164.0, 130.8, 126.1, 115.20, 69.3, 34.0,
33.0, 30.3, 30.2, 27.1, 23.7, 19.6, 14.4.
4-(nonyloxy)benzaldehyde (5-C9)
[0205] General procedure K was used to convert
4-hydroxybenzaldehyde (8.2 mmols) and 1-bromononane (12.3 mmols) to
6.5 mmols (79%) of the title product was recovered. .sup.1H NMR
(300 MHz, CDCl.sub.3) .delta. 9.88 (s, 1H), 7.83 (d, J=8.9 Hz, 2H),
6.99 (d, J=8.7 Hz, 2H), 4.04 (t, J=6.6 Hz, 2H), 1.76-1.85 (m, 2H),
1.14-1.53 (m, 12H), 0.88 (t, J=6.6 Hz, 3H); .sup.13C NMR (75 MHz,
CDCl.sub.3) .delta. 164.40, 132.11, 114.86, 68.55, 31.98, 29.47,
29.37, 29.18, 28.90, 28.31, 26.08, 22.79, 14.23.
4-(nonyloxy)benzoic acid (6-C9)
[0206] General procedure I was used to convert 5-C9 (1.5 g, 6.5
mmol) to 6-C9 (1.6 g, 6.5 mmol, 100%) .sup.1H NMR (300 MHz,
DMSO-d.sub.6) .delta. 7.87 (d, J=8.8 Hz, 2H), 6.99 (d, J=8.8 Hz,
2H), 4.02 (t, J=6.5 Hz, 2H), 1.71 (quin., J=6.8 Hz, 2H), 1.48-1.35
(m, 2H), 1.25 (bs, 12H), 0.85 (t, J=6.6 Hz, 3H); .sup.13C NMR (75
MHz, CDCl.sub.3) .delta. 167.0, 162.3, 131.3, 122.8, 114.2, 67.7,
31.3, 29.0, 28.7, 28.6, 28.5, 25.4, 22.1, 13.9.
N-(1-cyanocyclopropyl)-4-(nonyloxy)benzamide (7-C9)
[0207] General procedure G was used to convert 1.00 mmols of 6-C9
to an acid chloride. After standard purification techniques, the
acyl-chloride was isolated in quantitative yield and immediately.
General procedure F was used to couple 0.84 mmols of
1-amino-1-cyclopropanecarbonitrile. After standard purification
techniques, 0.59 mmols (70%) of the title product was recovered.
.sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 7.71 (d, J=8.9 Hz, 2H),
6.90 (d, J=8.9 Hz, 2H), 3.99 (t, J=6.6 Hz, 2H), 1.79 (quin., J=6
Hz, 1H), 1.61-1.65 (m, 2H), 1.19-1.50 (m, 14H), 0.88 (t, J=6.8 Hz,
1H); .sup.13C NMR (75 MHz, CDCl.sub.3) .delta. 129.4, 114.7, 95.6,
68.6, 38.0, 32.2, 29.9, 29.6, 29.4, 26.3, 23.0, 17.3, 14.5.
VPC171147
[0208] General procedure H was used to convert 0.20 mmols of 7-C9
to the title product. After standard purification techniques, 0.08
mmols (40%) of product was recovered and submitted for biological
evaluation. .sup.1H NMR (500 MHz, CD.sub.3OD) .delta. 7.88 (d,
J=8.6 Hz, 2H), 6.97 (d, J=8.6 Hz, 2H), 4.03 (t, J=6.3 Hz, 2H), 1.78
(m, 4H), 1.56 (bs, 2H), 1.47 (bd, 2H), 1.29 (bs, 8H), 0.90 (t,
J=6.7 Hz, 1H); .sup.13C NMR (125 MHz, CD.sub.3OD) .delta. 164.1,
130.8, 130.5, 126.1, 115.3, 115.2, 69.3, 33.1, 30.7, 30.5, 30.4,
30.3, 27.1, 23.8, 19.6, 17.0, 14.5; t.sub.R=10.60 min,
(m/z)=347.19.
4-(undecyloxy)benzaldehyde (5-C11)
[0209] General procedure K was used to convert
4-hydroxybenzaldehyde (8.2 mmols) and 1-bromononane (12.3 mmols) to
6.5 mmols (79%) of the title product was recovered. .sup.1H NMR
(300 MHz, CDCl.sub.3) .delta. 9.87 (s, 1H), 7.82 (d, J=8.8 Hz, 2H),
6.99 (d, J=8.8 Hz, 2H), 4.03 (t, J=6.5 Hz, 3H), 1.76-1.87 (m, 2H),
1.26-1.48 (m, 16H), 0.88 (t, J=6.7 Hz, 3H); .sup.13C NMR (75 MHz,
CDCl.sub.3) .delta. 191.0, 164.4, 132.1, 129.8, 114.9, 68.6, 34.3,
33.0, 32.1, 29.7, 29.6, 29.5, 29.2, 29.0, 28.3, 26.1, 22.8,
14.3.
4-(undecyloxy)benzoic acid (6-C11)
[0210] General procedure I was used to convert 5-C11 (1.5 g, 6.5
mmol) to 6-C11 (1.6 g, 6.5 mmol, 100%) .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta. 8.05 (d, J=8.8 Hz, 2H), 6.93 (d, J=8.8 Hz, 2H),
4.02 (t, J=6.6 Hz, 3H), 1.76-1.84 (m, 2H), 1.39-1.50 (m, 2H),
1.19-1.39 (m, 14H), 0.88 (t, J=6.8 Hz, 3H); .sup.13C NMR (75 MHz,
DMSO-d.sub.6) .delta. 167.0, 162.3, 131.3, 122.8, 114.2, 67.7,
31.3, 29.0, 28.7, 28.5, 25.4, 22.1, 13.9.
N-(1-cyanocyclopropyl)-4-(undecyloxy)benzamide (7-C11)
[0211] General procedure G was used to convert 1.00 mmols of 6-C1
to an acid chloride. After standard purification techniques, the
acyl-chloride was isolated in quantitative yield and immediately.
General procedure F was used to couple 0.84 mmols of
1-amino-1-cyclopropanecarbonitrile. After standard purification
techniques, 0.27 mmols (32%) of the title product was recovered.
.sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 7.71 (d, J=8.6 Hz, 2H),
6.90 (d, J=8.6 Hz, 2H), 3.98 (t, J=6.6 Hz, 2H), 1.79 (quin., J=5
Hz, 2H), 1.62 (dd, J=9.1, 6.3 Hz, 2H), 1.45 (quin., J=5 Hz, 2H),
1.35 (dd, J=9.0 Hz, 6.1 Hz, 2H), 1.20-1.32 (m, 14H), 0.88 (t, J=6.9
Hz, 3H); .sup.13C NMR (125 MHz, CDCl.sub.3) .delta. 168.2, 162.8,
129.6, 124.8, 120.8, 114.6, 68.5, 32.2, 29.90, 29.87, 29.68, 29.65,
29.4, 26.3, 23.0, 21.2, 17.2, 14.4.
VPC171167
[0212] General procedure H was used to convert 0.20 mmols of 7-C11
to the title product. After standard purification techniques, 0.07
mmols (35%) of product was recovered and submitted for biological
evaluation. .sup.1H NMR (500 MHz, CD.sub.3OD) .delta. 7.88 (d,
J=8.6 Hz, 2H), 6.97 (d, J=8.6 Hz, 2H), 4.03 (t, J=6.3 Hz, 2H),
1.81-1.74 (m, 4H), 1.56 (t, J=6.9 Hz, 2H), 1.47 (quin., J=6.3 Hz,
2H), 1.41-1.21 (m, 14H), 0.90 (t, J=6.7 Hz, 3H); .sup.13C NMR (125
MHz, CD.sub.3OD) .delta. 174.8, 171.7, 164.9, 131.68, 131.4, 126.9,
116.0, 70.1, 34.9, 33.9, 31.6, 31.4, 31.1, 28.0, 24.6, 20.4, 17.9,
15.3; t.sub.R=10.69 min, (m/z)=374.16.
4-(tridecyloxy)benzaldehyde (5-C13)
[0213] General procedure K was used to convert
4-hydroxybenzaldehyde (8.2 mmols) and 1-bromononane (12.3 mmols) to
7.3 mmols (87%) of the title product was recovered. .sup.1H NMR
(300 MHz, CDCl.sub.3) .delta. 9.88 (s, 1H), 7.83 (d, J=8.8 Hz, 2H),
6.99 (d, J=8.7 Hz, 2H), 4.04 (t, J=6.5 Hz, 2H), 1.76-1.88 (m, 2H),
1.52-1.08 (m, 20H), 0.88 (t, J=6.7 Hz, 3H).
4-(tridecyloxy)benzoic acid (6-C13)
[0214] General procedure I was used to convert 5-C31 (1.5 g, 7.3
mmol) to 6-C13 (1.6 g, 7.3 mmol, 100%) .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 7.87 (d, J=8.9 Hz, 2H), 6.98 (d, J=8.9 Hz, 2H),
4.01 (t, J=6.5 Hz, 2H), 1.80-1.71 (m, 2H), 1.49-1.13 (m, 20H), 0.85
(t, J=6.5 Hz, 3H); .sup.13C NMR (75 MHz, CDCl.sub.3) .delta. 166.9,
162.2, 131.3, 122.8, 114.1, 67.7, 35.1, 32.2, 31.3, 29.0, 28.9,
28.7, 28.5, 28.1, 27.5, 25.4, 22.1, 13.9.
N-(1-cyanocyclopropyl)-4-(tridecyloxy)benzamide (7-C13)
[0215] General procedure G was used to convert 1.00 mmols of 6-C13
to an acid chloride. After standard purification techniques, the
acyl-chloride was isolated in quantitative yield and immediately.
General procedure F was used to couple 0.84 mmols of
1-amino-1-cyclopropanecarbonitrile. After standard purification
techniques, 0.27 mmols (32%) of the title product was recovered.
.sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 7.72 (d, J=8.9 Hz, 2H),
6.90 (d, J=8.9 Hz, 2H), 3.98 (t, J=6.6 Hz, 2H), 1.86-1.69 (m, 2H),
1.61 (dd, J=10.0, 7.3 Hz, 2H), 1.51-1.01 (m, 22H), 0.87 (t, J=6.0
Hz, 3H); .sup.13C NMR (75 MHz, CDCl.sub.3) .delta. 129.4, 129.3,
114.7, 68.6, 68.5, 51.6, 32.3, 30.0, 29.7, 29.4, 26.3, 17.3.
VPC171137
[0216] General procedure H was used to convert 0.20 mmols of 7-C13
to the title product. After standard purification techniques, 0.06
mmols (30%) of product was recovered and submitted for biological
evaluation. .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. 9.16 (s,
1H), 7.80 (d, J=8.1 Hz, 2H), 6.99 (d, J=8.1 Hz, 2H), 4.01 (t, J=6.3
Hz, 2H), 1.70 (quin., J=5 Hz, 2H), 1.53 (t, J=6.5 Hz, 2H), 1.38
(quin., J=6.5 Hz, 2H), 1.27 (bs, 18H), 0.85 (t, J=6.5 Hz, 3H);
.sup.13C NMR (125 MHz, CD.sub.3OD) .delta. 167.7, 162.6, 130.3,
125.8, 122.1, 115.0, 68.7, 32.3, 30.0, 29.9, 29.7, 29.5, 26.4,
23.1, 21.3, 16.7, 14.9; t.sub.R=11.57 min, (m/z)=402.22.
N-(1-cyanocyclopropyl)palmitamide (8)
[0217] General procedure F was used to couple 1.68 mmols of
1-amino-1-cyclopropanecarbonitrile to 2.184 mmols of palmitoyl
chloride. After standard purification techniques, 1.3 mmols (77%)
of the title product was recovered. .sup.1H NMR (500 MHz, CDCl3)
.delta. 5.95 (s, 1H), 2.16 (t, J=7.6, 2H), 1.65-1.59 (m, 2H), 1.58
(s, 1H), 1.57-1.53 (m, 2H), 1.39-1.15 (m, 28H), 0.88 (dd, J=6.0,
7.0, 3H). .sup.13C NMR (126 MHz, CDCl.sub.3) .delta. 119.97, 36.08,
31.91, 29.64, 25.17, 22.68, 20.32, 16.85, 14.25.
VPC95291
[0218] General procedure H was used to convert 0.18 mmols of 8 to
the corresponding amidine. After standard purification techniques,
0.06 mmols (33%) of the title product were recovered. .sup.1H NMR
(500 MHz, CD.sub.3OD) .delta. 2.28-2.23 (m, 1H), 2.16 (t, J=7.5,
OH), 1.65 (dd, J=6.0, 8.3, 1H), 1.63-1.55 (m, 2H), 1.47 (dd, J=5.7,
8.2, OH), 1.38 (dd, J=6.0, 8.4, 1H), 1.29 (d, J=10.5, 19H), 1.18
(dd, J=5.6, 8.2, 1H), 0.90 (t, J=6.9, 3H). .sup.13C NMR (126 MHz,
CD.sub.3OD) .delta. 176.72, 172.34, 35.17, 32.02, 31.65, 29.35,
29.33, 29.30, 29.18, 29.05, 29.02, 28.95, 28.91, 28.74, 25.11,
24.79, 22.30, 17.95, 15.39, 13.00. t.sub.R=10.84 min.; m/z=339
N-(1-cyanocyclopropyl)oleamide (9)
[0219] General procedure F was used to 1.68 mmols of
1-amino-1-cyclopropanecarbonitrile to 2.184 mmols of oleoyl
chloride. After standard purification techniques, 1.24 mmols (74%)
of the title product was recovered. .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta. 6.53 (s, 1H), 5.43-5.25 (m, 2H), 2.17 (t,
J=7.6, 2H), 1.99 (dd, J=6.5, 12.5, 4H), 1.60 (d, J=7.2, 2H), 1.52
(dd, J=5.7, 8.3, 2H), 1.39-1.15 (m, 24H), 0.87 (t, J=6.9, 3H).
.sup.13C NMR (126 MHz, CDCl.sub.3) .delta. 174.09, 130.00, 129.66,
120.14, 97.26, 35.97, 31.88, 29.74, 29.68, 29.51, 29.30, 29.23,
29.14, 29.09, 27.20, 27.14, 25.22, 22.66, 20.39, 16.74, 14.11.
VPC95301
[0220] General procedure H was used to convert 0.44 mmols of 13 to
the title product. After standard purification techniques, 0.1
mmols (23%) was recovered and submitted for biological evaluation.
t.sub.R=11.13 min.; m/z=364.31
1-cyano-N-(4-decylphenyl)cyclopropanecarboxamide (10-C10)
[0221] General procedure E was used to convert 1.00 mmol of
4-decylanaline to title product; isolated as a white solid, 325 mg
(0.99 mmol, 99%): R.sub.f=0.52 (20% EtOAc in hexanes); .sup.1H NMR
(300 MHz, CDCl3) .delta. 8.16 (s, 1H), 7.41 (d, J=8.4, 2H), 7.14
(d, J=8.4, 2H), 2.73-2.40 (m, 2H), 1.77 (dd, J=4.4, 8.2, 2H),
1.68-1.51 (m, 4H), 1.29 (m, 14H), 0.90 (t, J=6.6, 3H); .sup.13C NMR
(75 MHz, CDCl3) .delta. 163.54, 140.27, 134.81, 129.13, 120.87,
120.27, 77.83, 77.40, 76.98, 35.63, 32.16, 31.69, 29.86, 29.75,
29.60, 29.49, 22.95, 18.44, 14.39, 14.32.
VPC143033
[0222] General procedure H was used to convert 0.99 mmol of 10-C10
to the title product; white solid isolated, 191 mg (0.51 mmol,
51%): .sup.1H NMR (500 MHz, DMSO) .delta. 9.69 (s, 1H), 9.50-8.33
(m, 4H), 7.47 (d, J=8.4, 2H), 7.10 (d, J=8.4, 2H), 2.50 (d, J=7.8,
2H), 1.66-1.34 (m, 6H), 1.22 (m, 14H), 0.83 (t, J=6.9, 3H);
.sup.13C NMR (126 MHz, DMSO) .delta. 168.27, 166.44, 138.41,
136.53, 128.64, 128.58, 121.33, 121.28, 34.98, 31.73, 31.44, 29.98,
29.45, 29.30, 29.13, 29.02, 22.54, 15.12, 14.41.
1-cyano-N-(4-undecylphenyl)cyclopropanecarboxamide (10-C11)
[0223] General procedure E was used to convert 1.00 mmol of
4-undecylanaline to title product; isolated as a white solid, 353
mg (0.99 mmol, 99%): R.sub.f=0.56 (20% EtOAc in hexanes); .sup.1H
NMR (300 MHz, CDCl3) .delta. 8.11 (s, 1H), 7.40 (d, J=8.4, 2H),
7.15 (d, J=8.4, 2H), 2.76-2.38 (m, 2H), 1.77 (dd, J=4.4, 8.2, 2H),
1.56 (m, J=4.4, 8.2, 4H), 1.27 (m, 18H), 0.89 (t, J=6.6, 3H);
.sup.13C NMR (75 MHz, CDCl3) .delta. 163.57, 140.34, 134.70,
129.20, 120.80, 120.28, 35.65, 32.19, 31.70, 29.92, 29.76, 29.63,
29.50, 22.95, 18.45, 14.34.
VPC143081
[0224] General procedure H was used to convert 0.99 mmol of 10-C11
to the title product; white solid isolated, 183 mg (0.39 mmol,
39%): .sup.1H NMR (500 MHz, DMSO) .delta. 9.69 (s, 1H), 9.50-8.33
(m, 4H), 7.45 (d, J=8.2, 2H), 7.12 (d, J=8.2, 2H), 2.50 (d, J=7.8,
2H), 1.66-1.34 (m, 6H), 1.22 (m, 18H), 0.83 (t, J=6.9, 3H);
.sup.13C NMR (126 MHz, DMSO) .delta. 168.22, 166.41, 138.46,
136.55, 128.69, 128.53, 121.33, 121.29, 34.98, 31.74, 31.47, 29.99,
29.45, 29.29, 29.13, 29.02, 22.54, 15.16, 14.49.
1-cyano-N-(4-dodecylphenyl)cyclopropanecarboxamide (10)
[0225] General procedure E was used to convert 1.00 mmol of
4-dodecylanaline to title product; isolated as a white solid, 353
mg (0.99 mmol, 99%): R.sub.f=0.59 (20% EtOAc in hexanes); .sup.1H
NMR (300 MHz, CDCl3) .delta. 8.11 (s, 1H), 7.40 (d, J=8.4, 2H),
7.15 (d, J=8.4, 2H), 2.76-2.38 (m, 2H), 1.77 (dd, J=4.4, 8.2, 2H),
1.56 (m, J=4.4, 8.2, 4H), 1.27 (m, 18H), 0.89 (t, J=6.6, 3H);
.sup.13C NMR (75 MHz, CDCl3) .delta. 163.57, 140.34, 134.70,
129.20, 120.80, 120.28, 35.65, 32.19, 31.70, 29.92, 29.76, 29.63,
29.50, 22.95, 18.45, 14.34.
VPC143064
[0226] General procedure H was used to convert 0.99 mmol of 10 to
the title product; white solid isolated, 183 mg (0.45 mmol, 45%):
.sup.1H NMR (500 MHz, DMSO) .delta. 9.69 (s, 1H), 9.50-8.33 (m,
4H), 7.45 (d, J=8.2, 2H), 7.12 (d, J=8.2, 2H), 2.50 (d, J=7.8, 2H),
1.66-1.34 (m, 6H), 1.22 (m, 18H), 0.83 (t, J=6.9, 3H); .sup.13C NMR
(126 MHz, DMSO) .delta. 168.22, 166.41, 138.46, 136.55, 128.69,
128.53, 121.33, 121.29, 34.98, 31.74, 31.47, 29.99, 29.45, 29.29,
29.13, 29.02, 22.54, 15.16, 14.49.
1-cyano-N-(4-tetradecylphenyl)cyclopropanecarboxamide (10-C14)
[0227] General procedure E was used to convert 1.00 mmol of
4-tetradecylanaline to title product; isolated as a white solid,
381 mg (0.99 mmol, 99%): R.sub.f=0.67 (20% EtOAc in hexanes);
.sup.1H NMR (300 MHz, CDCl3) .delta. 8.11 (s, 1H), 7.40 (d, J=8.4,
2H), 7.15 (d, J=8.4, 2H), 2.76-2.38 (m, 2H), 1.77 (dd, J=4.4, 8.2,
2H), 1.56 (dd, J=4.4, 8.2, 4H), 1.27 (m, 20H), 0.89 (t, J=6.6, 3H);
.sup.13C NMR (75 MHz, CDCl3) .delta. 163.54, 140.32, 134.67,
129.24, 120.82, 120.28, 35.62, 32.16, 31.71, 29.92, 29.73, 29.63,
29.54, 22.92, 18.44, 14.31.
VPC143065
[0228] General procedure H was used to convert 0.99 mmol of 10-C14
to the title product; white solid isolated, 161 mg (0.37 mmol,
37%): .sup.1H NMR (500 MHz, DMSO) .delta. 9.69 (s, 1H), 9.50-8.33
(m, 4H), 7.45 (d, J=8.2, 2H), 7.12 (d, J=8.2, 2H), 2.50 (d, J=7.8,
2H), 1.66-1.34 (m, 6H), 1.22 (m, 20H), 0.83 (t, J=6.9, 3H);
.sup.13C NMR (126 MHz, DMSO) .delta. 168.24, 166.43, 138.41,
136.52, 128.67, 128.51, 121.35, 121.29, 34.95, 31.74, 31.47, 29.97,
29.45, 29.30, 29.14, 29.02, 22.54, 15.16, 14.42.
1-cyano-N-(4-hexadecylphenyl)cyclonronanecarboxamide (10-C16)
[0229] General procedure E was used to convert 1.00 mmol of
4-hexadecylanaline to title product; isolated as a white solid, 409
mg (0.99 mmol, 99%): R.sub.f=0.85 (20% EtOAc in hexanes); .sup.1H
NMR (300 MHz, CDCl3) .delta. 8.14 (s, 1H), 7.40 (d, J=8.4, 2H),
7.14 (d, J=8.4, 2H), 2.78-2.44 (m, 2H), 1.77 (dd, J=4.4, 8.2, 2H),
1.56 (dd, J=4.4, 8.2, 4H), 1.27 (m, 22H), 0.89 (t, J=6.7, 3H);
.sup.13C NMR (75 MHz, CDCl3) .delta. 163.50, 140.29, 134.78,
129.14, 120.83, 120.27, 35.63, 32.19, 31.71, 29.95, 29.77, 29.64,
29.51, 22.96, 18.44, 14.40.
VPC143066
[0230] General procedure H was used to convert 0.99 mmol of 10-C16
to the title product; white solid isolated, 125 mg (0.27 mmol,
27%): .sup.1H NMR (500 MHz, DMSO) .delta. 9.69 (s, 1H), 9.50-8.33
(m, 4H), 7.45 (d, J=8.2, 2H), 7.12 (d, J=8.2, 2H), 2.50 (d, J=7.8,
2H), 1.66-1.34 (m, 6H), 1.22 (m, 22H), 0.83 (t, J=6.9, 3H);
.sup.13C NMR (126 MHz, DMSO) .delta. 168.25, 166.50, 138.51,
136.62, 128.69, 128.59, 121.41, 121.35, 34.99, 31.79, 31.41, 29.96,
29.45, 29.30, 29.12, 29.02, 22.54, 15.16, 14.39.
((oct-7-en-1-yloxy)methyl)cyclohexane (11)
[0231] General procedure J was used to convert 6.56 mmol of
7-bromo-1-heptene to 5.73 mmol (87%) of the title compound.
1-(7-(cyclohexylmethoxy)heptyl)-4-nitrobenzene (12)
[0232] General procedure M was used to convert 5.73 mmol of 11 and
6.30 mmol of 1-bromo-4-nitrobenzene to 3.72 mmol (65%) of the title
compound.
4-(7-(cyclohexylmethoxy)heptyl)aniline (13)
[0233] General procedure N was used to convert 0.657 mmol of 12 to
0.616 mmol (94%) of the title compound.
1-cyano-N-(4-(7-(cyclohexylmethoxy)heptyl)phenyl)cyclopropanecarboxamide
(14)
[0234] General procedure E was used to convert 0.616 mmol of 13 and
0.924 mmol of 1-cyanocyclopropanecarboxylic acid to 0.590 mmol
(96%) of the title compound.
VPC143105
[0235] General procedure H was used to convert 0.590 mmol of 14 to
0.349 mmol (59%) of the title product and submitted for biological
evaluation.
((hept-6-en-1-yloxy)methyl)benzene (11a)
[0236] General procedure J was used to convert 6.56 mmol of
7-bromo-1-heptene to 5.90 mmol (90%) of the title compound.
1-(7-(benzyloxy)heptyl)-4-nitrobenzene (12a)
[0237] General procedure M was used to convert 5.90 mmol of 11a and
6.49 mmol of 1-bromo-4-nitrobenzene to 4.03 mmol (68%) of the title
compound.
4-(7-(benzyloxy)heptyl)aniline (13a)
[0238] General procedure N was used to convert 1.10 mmol of 12a to
1.05 mmol (95%) of the title compound.
N-(4-(7-(benzyloxy)heptyl)phenyl)-1-cyanocyclopropanecarboxamide
(14a)
[0239] General procedure E was used to convert 1.05 mmol of 13a and
1.575 mmol of 1-cyanocyclopropanecarboxylic acid to 1.03 mmol (98%)
of the title compound.
VPC143078
[0240] General procedure H was used to convert 1.03 mmol of 14a to
0.330 mmol (32%) of the title product and submitted for biological
evaluation.
((hex-5-en-1-yloxy)methyl)cyclohexane (11b)
[0241] General procedure J was used to convert 7.48 mmol of
6-bromo-1-hexene to 5.21 mmol (70%) of the title compound.
1-(6-(cyclohexylmethoxy)hexyl)-4-nitrobenzene (12b)
[0242] General procedure M was used to convert 5.21 mmol of 11b and
5.73 mmol of 1-bromo-4-nitrobenzene to 1.35 mmol (26%) of the title
compound.
4-(6-(cyclohexylmethoxy)hexyl)aniline (13b)
[0243] General procedure N was used to convert 1.35 mmol of 12b to
1.30 mmol (96%) of the title compound.
1-cyano-N-(4-(6-(cyclohexylmethoxy)hexyl)phenyl)cclopropanecarboxamide
(14b)
[0244] General procedure E was used to convert 1.30 mmol of 13b and
1.95 mmol of 1-cyanocyclopropanecarboxylic acid to 1.29 mmol (99%)
of the title compound.
VPC143113
[0245] General procedure H was used to convert 1.29 mmol of 14b to
0.581 mmol (45%) of the title product and submitted for biological
evaluation.
((oct-7-en-1-yloxy)methyl)cyclohexane (11c)
[0246] General procedure J was used to convert 5.98 mmol of
8-bromo-1-octene to 4.56 mmol (77%) of the title compound.
1-(8-(cyclohexylmethoxy)octyl)-4-nitrobenzene (12c)
[0247] General procedure M was used to convert 4.56 mmol of 11c and
5.02 mmol of 1-bromo-4-nitrobenzene to 2.19 mmol (48%) of the title
compound.
4-(8-(cyclohexylmethoxy)octyl)aniline (13c)
[0248] General procedure N was used to convert 2.19 mmol of 12c to
2.05 mmol (94%) of the title compound.
1-cyano-N-(4-(8-(cyclohexylmethoxy)octyl)phenyl)cyclopropanecarboxamide
(14c)
[0249] General procedure E was used to convert 2.05 mmol of 13c and
3.08 mmol of 1-cyanocyclopropanecarboxylic acid to 2.01 mmol (98%)
of the title compound.
VPC143126
[0250] General procedure H was used to convert 2.01 mmol of 14c to
0.679 mmol (34%) of the title product and submitted for biological
evaluation.
2-((hept-6-en-1-yloxy)methyl)adamantane (11d)
[0251] General procedure J was used to convert 3.26 mmol of
7-bromo-1-heptene to 2.77 mmol (85%) of the title compound.
2-(((7-(4-nitrophenyl)heptyl)oxy)methyl)adamantane (12d)
[0252] General procedure M was used to convert 2.77 mmol of 11d and
3.05 mmol of 1-bromo-4-nitrobenzene to 1.30 mmol (47%) of the title
compound.
4-(7-(adamantan-2-ylmethoxy)heptyl)aniline (13d)
[0253] General procedure N was used to convert 1.30 mmol of 12d to
1.30 mmol (100%) of the title compound.
N-(4-(7-(adamantan-2-ylmethoxy)heptyl)phenyl)-1-cyanocyclopropanecarboxami-
de (14d)
[0254] General procedure E was used to convert 1.30 mmol of 13d and
1.95 mmol of 1-cyanocyclopropanecarboxylic acid to 1.25 mmol (96%)
of the title compound.
VPC143090
[0255] General procedure H was used to convert 1.25 mmol of 14d to
0.891 mmol (71%) of the title product and submitted for biological
evaluation.
non-8-en-1-ylbenzene (15)
[0256] General procedure L was used to convert 5.98 mmol of
8-bromo-1-octene to 5.08 mmol (85%) of the title compound.
1-nitro-4-(9-phenylnonyl)benzene (16)
[0257] General procedure M was used to convert 5.08 mmol of 15 and
5.59 mmol of 1-bromo-4-nitrobenzene to 2.74 mmol (54%) of the title
compound.
4-(9-phenylnonyl)aniline (17)
[0258] General procedure N was used to convert 2.74 mmol of 16 to
2.56 mmol (93%) of the title compound.
1-cyano-N-(4-(9-phenylnonyl)phenyl)cyclopropanecarboxamide (18)
[0259] General procedure E was used to convert 2.56 mmol of 17 and
3.84 mmol of 1-cyanocyclopropanecarboxylic acid to 2.40 mmol (94%)
of the title compound.
VPC143129
[0260] General procedure H was used to convert 2.40 mmol of 18 to
1.12 mmol (47%) of the title product and submitted for biological
evaluation.
oct-7-en-1-ylbenzene (15a)
[0261] General procedure L was used to convert 6.52 mmol of
7-bromo-1-heptene to 5.45 mmol (84%) of the title compound.
1-nitro-4-(8-phenyloctyl)benzene (16a)
[0262] General procedure M was used to convert 5.31 mmol of 15a and
5.84 mmol of 1-bromo-4-nitrobenzene to 3.42 mmol (71%) of the title
compound.
4-(8-phenyloctyl)aniline (17a)
[0263] General procedure N was used to convert 3.42 mmol of 16a to
3.15 mmol (92%) of the title compound.
1-cyano-N-(4-(8-phenyloctyl)phenyl)cyclopropanecarboxamide
(18a)
[0264] General procedure E was used to convert 3.15 mmol of 17a and
4.73 mmol of 1-cyanocyclopropanecarboxylic acid to 3.10 mmol (98%)
of the title compound.
VPC143119
[0265] General procedure H was used to convert 3.10 mmol of 18a to
1.34 mmol (43%) of the title product and submitted for biological
evaluation.
N-(1-cyanocyclopropyl)-4-(7-(cyclohexylmethoxy)heptyl)benzamide
(21)
[0266] General procedure G was used to convert 1.89 mmols of 20 to
an acid chloride. After standard purification techniques, the
acyl-chloride was isolated in quantitative yield and immediately.
General procedure F was used to couple 1.89 mmols of
1-amino-1-cyclopropanecarbonitrile. After standard purification
techniques, 1.34 mmols (71%) of the title product was recovered
VPC14a1051
[0267] General procedure H was used to convert 1.34 mmol of 21 to
1.13 mmol (84%) of the title product and submitted for biological
evaluation.
((hept-6-en-1-yloxy)methyl)cyclopentane (11e)
[0268] General procedure J was used to convert 3.26 mmol of
7-bromo-1-heptene to 2.86 mmol (88%) of the title compound.
4-(7-(cyclopentylmethoxy)heptyl)benzaldehyde (19a)
[0269] General procedure M was used to convert 2.86 mmol of 11e and
3.15 mmol of 1-bromo-4-nitrobenzene to 2.29 mmol (80%) of the title
compound.
4-(7-(cyclopentylmethoxy)heptyl)benzoic acid (20a)
[0270] General procedure I was used to convert 3.15 mmol of 19a to
3.15 mmol (100%) of the title compound.
N-(1-cyanocyclopropyl)-4-(7-(cyclopentylmethoxy)heptyl)benzamide
(21a)
[0271] General procedure G was used to convert 3.15 mmols of 20a to
an acid chloride. After standard purification techniques, the
acyl-chloride was isolated in quantitative yield and immediately.
General procedure F was used to couple 3.15 mmols of
1-amino-1-cyclopropanecarbonitrile. After standard purification
techniques, 2.05 mmols (65%) of the title product was
recovered.
VPC143154
[0272] General procedure H was used to convert 2.05 mmol of 21a to
1.76 mmol (86%) of the title product and submitted for biological
evaluation.
((dec-9-en-1-yloxy)methyl)cyclohexane (22)
[0273] General procedure J was used to convert 4.90 mmol of
10-bromo-1-decene to 3.62 mmol (74%) of the title compound.
10-(cyclohexylmethoxy)decan-1-ol (23)
[0274] General procedure O was used to convert 3.62 mmol of 22 to
2.72 mmol (75%) of the title compound. .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 4.33 (d, J=2.8 Hz, 1H), 3.53 (t, J=6.6 Hz, 2H),
3.30 (t, J=6.7 Hz, 2H), 3.12 (d, J=6.6 Hz, 2H), 2.18 (s, 1H), 1.52
(m, 16H), 1.20 (m, 8H), 0.83 (m, 2H). .sup.13C NMR (75 MHz,
CDCl.sub.3) .delta. 76.90, 71.61, 71.18, 70.76, 69.57, 68.06,
64.21, 62.23, 60.28, 41.95, 38.33, 37.33, 34.66, 34.09, 33.53,
32.99, 32.48, 31.76, 30.35, 29.85, 27.76, 26.92, 26.74, 26.46,
26.15, 25.44, 22.98, 22.33, 20.75, 19.35, 14.26, 13.79.
(((10-azidodecyl)oxy)methyl)cyclohexane (24)
[0275] General procedure P was used to convert 2.72 mmol of 23 to
2.07 mmol (76%) of the title compound. .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 3.31 (t, J=6.6 Hz, 2H), 3.13 (d, J=6.5 Hz, 2H),
1.66 (m, 17H), 1.21 (m, 12H). .sup.13C NMR (75 MHz, CDCl.sub.3)
.delta. 71.25, 69.78, 62.33, 58.18, 51.61, 38.26, 37.25, 33.85,
30.95, 30.82, 30.36, 29.93, 29.85, 29.63, 29.33, 29.03, 27.41,
26.89, 26.56, 26.37, 26.10, 25.86, 25.29, 23.33, 21.01, 14.34.
1-cyano-N-(10-(cyclohexylmethoxy)decyl)cyclopropanecarboxamide
(26)
[0276] General procedure Q was used to convert 2.72 mmol of 24 to
0.626 mmol (23%) of the title compound. .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 6.61 (s, 1H) 3.33 (t, J=6.6 Hz, 2H), 3.23 (dd,
J=13.4, 6.8 Hz, 2H), 3.14 (d, J=6.6 Hz, 2H), 1.54 (m, 17H), 1.19
(m, 11H). 0.83 (m, 4H). .sup.13C NMR (75 MHz, CDCl.sub.3) .delta.
165.21, 120.49, 71.28, 50.94, 40.85, 38.23, 32.44, 30.91, 30.37,
29.93, 29.63, 29.41, 27.17, 26.98, 26.88, 26.36, 26.10, 25.67,
23.87, 17.63, 13.64.
VPC143144
[0277] General procedure H was used to convert 0.626 mmol of 26 to
0.134 mmol (21%) of the title product and submitted for biological
evaluation.
((dec-9-en-1-yloxy)methyl)cyclopentane (22a)
[0278] General procedure J was used to convert 4.90 mmol of
10-bromo-1-decene to 3.54 mmol (72%) of the title compound. .sup.1H
NMR (300 MHz, CDCl3) .delta. 5.78 (ddt, J=6.7, 10.2, 16.9, 1H),
4.92 (m, 2H), 3.37 (m, 4H), 2.52 (dq, J=7.2, 14.2, 1H), 2.02 (m,
3H), 1.85 (m, 2H), 1.70 (m, 2H), 1.53 (m, 3H), 1.30 (d, J=14.9,
8H). .sup.13C NMR (75 MHz, CDCl.sub.3) .delta. 139.12, 114.27,
71.24, 35.42, 34.00, 29.93, 29.65, 29.28, 29.11, 26.36, 25.30,
18.78.
10-(cyclopentylmethoxy)decan-1-ol (23a)
[0279] General procedure O was used to convert 3.54 mmol of 22a to
2.59 mmol (73%) of the title compound. .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta. 3.58 (t, J=6.7, 2H), 3.36 (m, 4H), 2.52 (dq,
J=7.4, 14.8, 1H), 2.02 (m, 3H), 1.83 (m, 2H), 1.65 (m, 3H), 1.49
(m, 6H), 1.23 (dd, J=6.5, 13.9, 8H). .sup.13C NMR (126 MHz,
CDCl.sub.3) .delta. 75.15, 70.77, 70.09, 59.99, 38.16, 36.81,
35.05, 31.83, 29.27, 28.05, 26.86, 25.22, 22.07, 20.62, 18.38,
13.94.
(((10-azidodecyl)oxy)methyl)cyclopentane (24a)
[0280] General procedure P was used to convert 2.59 mmol of 23a to
1.87 mmol (72%) of the title compound. .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 3.35 (t, J=7.0, 4H), 3.20 (t, J=6.9, 2H), 2.52
(s, 1H), 1.99 (s, 3H), 1.84 (d, J=6.0, 3H), 1.69 (d, J=9.8, 3H),
1.52 (s, 5H), 1.25 (s, 8H). .sup.13C NMR (75 MHz, CDCl.sub.3)
.delta. 77.82, 71.24, 51.60, 35.39, 30.92, 29.90, 29.63, 29.33,
29.03, 27.41, 26.89, 26.33, 25.33, 23.31, 18.80.
1-cyano-N-(10-(cyclopentylmethoxy)decyl)cyclopropanecarboxamide
(26a)
[0281] General procedure Q was used to convert 1.87 mmol of 24a to
0.511 mmol (27%) of the title compound. .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 6.63 (s, 1H), 3.36 (t, J=6.8 Hz, 4H), 3.24 (tt,
J=18.4, 9.2 Hz, 2H), 2.51 (dq, J=14.4, 7.2 Hz, 1H), 2.01 (m, 2H),
1.84 (m, 2H), 1.58 (m, 14H), 1.25 (s, 8H). .sup.13C NMR (75 MHz,
CDCl.sub.3) .delta. 165.24, 120.52, 75.74, 71.29, 53.77, 40.86,
35.37, 29.90, 29.62, 29.40, 26.98, 26.32, 25.38, 19.28, 18.82,
17.64, 13.66.
VPC14a1058
[0282] General procedure H was used to convert 0.511 mmol of 26a to
0.237 mmol (46%) of the title product and submitted for biological
evaluation.
1-cyano-N-tetradecylcyclopropanecarboxamide (27)
[0283] General procedure E was used; white solid isolated, 304 mg
(0.99 mmol, 99%): R.sub.f=0.45 (20% EtOAc in hexanes); .sup.1H NMR
(300 MHz, CDCl3) .delta. 6.41 (s, 1H), 3.46-3.15 (m, 2H), 1.66 (dd,
J=4.4, 8.1, 2H), 1.58-1.39 (m, 4H), 1.26 (m, 22H), 0.86 (t, J=6.7,
3H); .sup.13C NMR (75 MHz, CDCl3) .delta. 165.22, 120.54, 40.89,
32.14, 29.87, 29.79, 29.71, 29.58, 29.45, 27.01, 22.91, 17.65,
14.34, 13.67.
VPC143057
[0284] General procedure H was used; white solid isolated, 150 mg
(0.42 mmol, 42%): .sup.1H NMR (300 MHz, DMSO) .delta. 8.96 (s, 4H),
7.82 (s, 1H), 3.02 (s, 2H), 1.28 (m, 22H), 0.83 (s, 3H); .sup.13C
NMR (75 MHz, DMSO) .delta. 168.55, 167.92, 31.98, 29.74, 29.56,
29.48, 29.40, 29.09, 27.04, 22.78, 14.86, 14.64.
1-cyano-N-hexadecylcyclopropanecarboxamide (27a)
[0285] General procedure E was used; white solid isolated, 319 mg
(0.99 mmol, 99%): R.sub.f=0.47 (20% EtOAc in hexanes); .sup.1H NMR
(500 MHz, CDCl3) .delta. 6.37 (s, 1H), 3.28 (dd, J=6.6, 13.7, 2H),
1.73-1.60 (m, 2H), 1.53 (d, J=7.3, 2H), 1.47 (dd, J=4.3, 8.1, 1H),
1.27 (m, 24H), 0.87 (t, J=6.9, 3H); .sup.13C NMR (126 MHz, CDCl3)
.delta. 164.99, 120.33, 40.68, 31.91, 29.67, 29.56, 29.48, 29.38,
29.22, 26.78, 22.68, 17.43, 14.12.
VPC143046
[0286] General procedure H was used; white solid isolated, 174 mg
(0.45 mmol, 45%): .sup.1H NMR (300 MHz, DMSO) .delta. 8.96 (s, 4H),
7.82 (s, 1H), 3.02 (s, 2H), 1.28 (m, 24H), 0.83 (s, 3H); .sup.13C
NMR (126 MHz, DMSO) .delta. 168.25, 166.50, 138.51, 136.62, 128.69,
128.59, 121.41, 121.35, 34.99, 31.79, 31.41, 29.96, 29.45, 29.30,
29.12, 29.02, 22.54, 15.16, 14.39.
1-cyano-N-octadecylcyclopronanecarboxamide (27b)
[0287] General procedure E was used; white solid isolated (0.99
mmol, 99%): R.sub.f=0.47 (20% EtOAc in hexanes).
VPC14a1002
[0288] General procedure H was used; white solid isolated (0.05
mmol, 5%).
(S)-tert-butyl 2-cyanopyrrolidine-1-carboxylate (28)
[0289] To a flame dried flask was added 2 grams (9.3 mmols) of
N-Boc proline followed by 93 mL of dichloromethane and 4.24 grams
(41.85 mmols) of triethylamine. The reaction mixture was then
cooled to -25.degree. C. and 2.5 grams (18.6 mmols) of
iso-butylchloroformate was added dropwise. After stirring for 45
minutes at this low temperature, 0.83 grams (49 mmol) of ammonia
was added to the reaction mixture as a 7N solution in methanol. The
flask was allowed to warm to ambient temperature and stir for an
additional 16 hours. After this time, the reaction mixture was
evaporated to dryness and reconstituted in 200 mL of EtOAc. The
organic layer was washed four 15 mL portions of 1N HCl, dried with
MgSO.sub.4 and evaporated to dryness. The crude material was then
immediately dehydrated to the nitrile according to general
procedure E using 1.9 grams (18.88 mmols) of triethylamine and 2.34
grams (11.16 mmols) of trifluoroacetic anhydride in 93 mL of THF.
After standard work-up and purification techniques (column
chromatography, 25% EtOAc in hexanes), 0.91 grams (4.65 mmols, 50%)
of the title product was recovered as a yellow oil. .sup.1H NMR
(300 MHz, CDCl.sub.3) .delta. 4.53 (d, J=5.5 Hz, 0.5H), 4.48-4.38
(m, 0.5H), 3.57-3.41 (m, 1H), 3.41-3.22 (m, 1H), 2.31-1.92 (m, 4H),
1.47 (d, J=9.4 Hz, 9H). .sup.13C NMR (75 MHz, CDCl.sub.3) .delta.
153.25, 119.38, 81.64, 47.39, 45.93, 31.86, 31.01, 28.52, 24.88,
24.04.
(S)-1-(4-dodecylbenzoyl)pyrrolidine-2-carbonitrile (29)
[0290] General procedure D was used to deprotect 0.4 grams (2.05
mmols) of 28 using 21 mL of dichloromethane and trifluoroacetic
acid. After standard work-up procedures, the TFA salt of 28 was
coupled to 0.6 grams (2.05 mmols) of 4-dodecylbenzoic acid using
1.1 grams (2.05 mmols) of PyBOP and 1.1 grams (8.2 mmols) of
N,N-diisopropylethylamine in 21 mL of dichloromethane according
general procedure E and heating to 50.degree. C. After previously
described work-up and purification procedures (column
chromatography, 25% EtOAc in hexanes), 0.17 grams (0.45 mmols, 22%)
of the title product was recovered as an off-white solid. .sup.1H
NMR (500 MHz, CDCl.sub.3) .delta. 7.50 (d, J=8.0 Hz, 2H), 7.22 (d,
J=7.6 Hz, 2H), 4.96-4.84 (m, 1H), 3.73-3.63 (m, 1H), 5.62-3.52 (m,
1H), 2.68-2.58 (m, 2H), 2.39-2.26 (m, 1H), 2.24-2.10 (m, 1H),
2.06-1.96 (m, 1H), 1.67-1.55 (m, 1H), 1.39-1.18 (m, 18H), 0.87 (t,
J=6.9 Hz, 3H). .sup.13C NMR (126 MHz, CDCl.sub.3) .delta. 170.18,
146.28, 128.40, 127.61, 127.17, 118.73, 49.51, 46.87, 35.84, 31.91,
31.20, 30.30, 29.65, 29.56, 29.46, 29.34, 29.23, 25.58, 22.68,
14.12.
VPC96091
[0291] General procedure H was used to convert 0.21 grams (0.56
mmols) of 29 to the title product using 112 .mu.L of 0.5M sodium
methoxide in methanol (0.003 grams, 0.056 mmols), 6 mL of anhydrous
methanol and 0.033 grams (0.62 mmols) of ammonium chloride. After
previously described recrystallization procedures 0.084 grams (0.2
mmols, 36%) of VPC96091 was recovered as a powdery white solid and
submitted for biological evaluation. .sup.1H NMR (500 MHz,
CD.sub.3OD) .delta. 7.63 (d, J=8.0 Hz, 2H), 7.30 (d, J=8.0 Hz, 2H),
4.73 (t, J=7.5 Hz, 1H), 3.87 (dd, J=15.4, 9.0 Hz, 1H), 3.73-3.60
(m, 1H), 2.67 (t, J=7.6 Hz, 2H), 2.54 (dd, J=13.3, 6.6 Hz, 1H),
2.12-1.93 (m, 2H), 1.71-1.56 (m, 1H), 1.34-1.28 (m, 18H), 0.90 (t,
J=6.9 Hz, 3H). .sup.13C NMR (126 MHz, CD.sub.3OD) .delta. 171.79,
146.52, 131.91, 128.06, 127.63, 58.94, 50.59, 35.36, 31.66, 31.14,
31.03, 29.33, 29.28, 29.15, 29.06, 28.88, 25.27, 22.32, 13.03.
(S)-1-(4-undecylbenzoyl)pyrrolidine-2-carbonitrile (29a)
[0292] General procedure D was used to deprotect 0.447 grams (2.3
mmols) of 28. After standard work-up procedures, general procedure
E was used to couple 2.3 mmols of the TFA salt of 28 to 0.635 grams
(2.3 mmols) of 4-undecylbenzoic acid at 50.degree. C. After
work-up, 0.334 grams (0.94 mmols, 41%) of the title product was
isolated as a yellow solid by flash chromatography (25% EtOAc in
hexanes). .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 7.24 (dd,
J=12.2, 4.4 Hz, 2H), 7.04 (t, J=7.7 Hz, 2H), 4.97 (s, 1H), 3.88 (s,
1H), 3.31 (dt, J=18.1, 9.0 Hz, 1H), 2.77-2.64 (m, 1H), 2.63-2.54
(m, 1H), 2.24-2.12 (m, 1H), 1.60 (dt, J=15.1, 7.3 Hz, 4H), 1.29 (d,
J=10.9 Hz, 14H), 0.88 (t, J=6.7 Hz, 3H). .sup.13C NMR (126 MHz,
CDCl.sub.3) .delta. 169.97, 146.29, 128.42, 128.18, 127.63, 127.17,
118.73, 49.51, 46.85, 46.20, 35.84, 31.91, 31.20, 30.27, 29.64,
29.61, 29.56, 29.46, 29.33, 29.23, 25.57, 22.68, 14.12.
VPC96077
[0293] General procedure H was used to convert 0.148 grams (0.42
mmols) of 29a to the title product. After standard
recrystallization techniques, 0.068 grams (0.16 mmols, 38%) of the
amidine was recovered as a powdery white solid.
(S)-1-(4-octylbenzoyl)pyrrolidine-2-carbonitrile (29b)
[0294] General procedure D was used to deprotect 0.4 grams (2.05
mmols) of 28 in 21 mL of dichloromethane and trifluoroacetic acid.
After work-up, the TFA salt of 28 was then coupled to 0.6 grams
(2.05 mmols) of 4-octylbenzoic acid with 1.1 grams (2.05 mmols) of
PyBOP and 1.1 grams (8.2 mmols) of N,N-diisopropylethylamine in 21
mL of dichloromethane according to general procedure E and heating
at 50.degree. C. After previously described work-up and
purification procedures (column chromatography, 25% EtOAc in
hexanes) 0.256 grams (0.82 mmols, 40%) of the title product was
recovered.
VPC95127
[0295] General procedure H was used to convert 0.1 grams (0.32
mmols) of (S)-1-(4-octylbenzoyl)pyrrolidine-2-carbonitrile to the
title product using 0.032 mmols of sodium methoxide as a 0.5 M
solution in methanol and 0.019 grams (0.352 mmols) of ammonium
chloride. After standard work-up procedures, 0.039 grams (0.107
mmols, 33%) of the title product was recovered as an off white
solid. .sup.1H NMR (500 MHz, CD.sub.3OD) .delta. 7.62 (d, J=5.6 Hz,
2H), 7.31 (d, J=7.8 Hz, 2H), 4.71 (dd, J=6.9, 4.4 Hz, 2H), 3.85 (s,
1H), 3.68 (s, 1H), 2.67 (t, J=7.8 Hz, 2H), 2.54 (d, J=9.1 Hz, 1H),
2.03 (d, J=24.3 Hz, 2H), 1.62 (d, J=13.3 Hz, 1H), 1.30 (m, 10H),
0.89 (t, J=6.7 Hz, 3H).
4-(octyloxy)benzaldehyde (30)
[0296] 4-hydroxybenzaldehyde (8.2 mmols), 1-bromooctane (12.3
mmols) and cesium carbonate (16.4 mmols) were dissolved in DMF
(13.7 mmols). The mixture was then heated to 80.degree. C. and
stirred for 3 hours. After this time, the reaction mixture was
filtered through a fine fritted funnel; the eluent was collected
and diluted with 200 mLs of EtOAc. The organic layer was extracted
with seven 10 mL portions of H.sub.2O and two 15 mL portions of
brine. The organic layer was dried with MgSO.sub.4 and evaporated
to dryness. After purification by flash chromatography 7.8 mmols of
the title product was recovered. .sup.1H NMR (500 MHz, CDCl.sub.3)
.delta. 9.86 (d, J=2.9, 1H), 7.94-7.74 (m, 2H), 7.05-6.92 (m, 2H),
4.02 (td, J=3.1, 6.5, 2H), 1.88-1.73 (m, 2H), 1.44 (dd, J=6.8,
14.2, 2H), 1.39-1.21 (m, 8H), 0.99-0.81 (m, 3H). .sup.13C NMR (126
MHz, CDCl.sub.3) .delta. 190.76, 164.24, 131.95, 129.70, 114.71,
68.40, 31.77, 29.28, 29.03, 25.95, 22.63, 14.08.
4-(octyloxy)benzoic acid (31)
[0297] General procedure I was used to convert 7.8 mmols of 30 to
the title product. After standard purification techniques, 7.8
mmols of the acid were recovered. .sup.1H NMR (500 MHz, CDCl.sub.3)
.delta. 8.06 (d, J=8.8, 2H), 6.93 (dd, J=2.2, 9.3, 2H), 4.02 (t,
J=6.6, 2H), 1.86-1.75 (m, 2H), 1.51-1.42 (m, 2H), 1.33 (ddd, J=7.5,
13.7, 18.7, 8H), 0.89 (t, J=6.9, 3H). .sup.13C NMR (126 MHz,
CDCl.sub.3) .delta. 171.96, 163.68, 132.32, 121.33, 114.17, 68.28,
31.78, 29.31, 29.07, 25.97, 22.64, 14.08.
(S)--N-(1-cyanoethyl)-4-(octyloxy)benzamide (32)
[0298] General procedure D and E were used to deprotect 1.17 mmols
of (S)-tert-butyl 1-cyanoethylcarbamate and couple the resulting
amine to 1.17 mmols of 31. After standard purification techniques
0.35 mmols of the title product were recovered. .sup.1H NMR (500
MHz, CDCl.sub.3) .delta. 7.74 (d, J=8.9, 2H), 6.90 (d, J=8.8, 2H),
6.67 (s, 1H), 5.30-5.05 (m, 1H), 3.98 (t, J=6.6, 2H), 1.85-1.74 (m,
2H), 1.64 (d, J=7.2, 3H), 1.45 (s, 3H), 1.30 (d, J=19.1, 7H), 0.88
(t, J=6.9, 3H). .sup.13C NMR (126 MHz, CDCl.sub.3) .delta. 166.26,
162.49, 129.11, 124.52, 119.58, 114.40, 68.26, 36.25, 31.77, 29.30,
29.19, 29.07, 25.95, 22.63, 19.53, 14.08.
VPC95229
[0299] General procedure H was used to convert 0.35 mmols of 32 to
the title product. After standard purification techniques, 1 mg of
the title product was recovered and submitted for biological
evaluation. t.sub.R=7.9 min.; m/z=320.37.
3-(benzyloxy)benzaldehyde (33)
[0300] General procedure K was used to convert
3-hydroxybenzaldehyde (8.2 mmols) and benzyl bromide (12.3 mmols)
to 1 (8.2 mmols, quant.) .sup.1H NMR (500 MHz, CDCl.sub.3) .delta.
9.98 (s, 1H), 7.51-7.44 (m, 4H), 7.44-7.39 (m, 1H), 7.38-7.33 (m,
1H), 7.26 (ddd, J=1.8, 3.5, 4.2, 1H), 5.13 (s, 2H). .sup.13C NMR
(126 MHz, CDCl.sub.3) .delta. 192.08, 159.29, 137.81, 136.29,
130.13, 128.68, 128.22, 127.56, 123.70, 122.20, 113.22, 77.31,
77.06, 76.80, 70.20.
(3-(benzyloxy)phenyl)methanol (34)
[0301] The intermediate 33 (8.2 mmols) and calcium chloride (16.4
mmols) were dissolved in 55 mL of anhydrous ethanol and 27 mLs of
THF. The solution was next cooled to 0.degree. C. and sodium
borohydride (16.4 mmols) was added slowly over several minutes to
the reaction vessel. Upon warming to ambient temperature, the
mixture was allowed to stir for an additional 15 hours. After this
time, the flask was again cooled to 0.degree. C. and the excess
sodium borohydride was quenched with the addition of 5 mLs of 1N
HCl. The aqueous layer was extracted with four 20 mL portions of
EtOAc; the organic layers were then combined, washed with 10 mLs of
brine and dried with MgSO.sub.4. After evaporation, the crude
organic material was purified by flash chromatography and 6.7 mmols
of the title product was recovered. .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 7.49-7.24 (m, 5H), 7.06-6.88 (m, 2H), 5.08 (s,
2H), 4.66 (s, 2H), 1.78 (s, 1H). .sup.13C NMR (75 MHz, CDCl.sub.3)
.delta. 129.88, 128.83, 128.21, 127.72, 119.60, 114.35, 113.45,
70.17, 65.46.
4-(3-(benzyloxy)benzyloxy)benzaldehyde (35)
[0302] To a stirring solution of triphenylphosphine (1.1 mmols) in
THF (2 mLs) was added 34 (1.1 mmols). The solution was immediately
cooled to 0.degree. C. and diisopropyl azodicarboxylate (1.1 mmols)
was added dropwise and the mixture was stirred at this low
temperature for 15 minutes. After this time, 4-hydroxybenzaldehyde
(1 mmol) was added to the reaction vessel, still stirring at
0.degree. C. The mixture was warmed slowly to ambient temperature
and stirred for 15 hours. At this time, the THF was evaporated,
reconstituted in Et.sub.2O and filtered through a small portion of
silica gel. The eluent was collected and evaporated to dryness. The
title product (0.45 mmols) was recovered after flash
chromatography. .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 9.89 (s,
1H), 7.91-7.77 (m, 2H), 7.50-7.30 (m, 6H), 7.03 (ddd, J=5.1, 11.5,
24.0, 5H), 5.13 (s, 2H), 5.08 (s, 2H). .sup.13C NMR (75 MHz,
CDCl.sub.3) .delta. 191.04, 163.88, 159.36, 137.82, 136.98, 132.24,
130.09, 128.86, 128.29, 127.74, 120.05, 115.39, 114.79, 114.14,
77.70, 77.27, 76.85, 70.25.
4-(3-(benzyloxy)benzyloxy)benzoic acid (36)
[0303] General procedure I was used to oxidize 0.45 mmols of 35 to
the corresponding carboxylic acid. The title product was recovered
in quantitative yield; no further purification was carried out
other than that outlined in the general procedure. The recovered
product was extremely insoluble in deuterated NMR solvents and no
spectra could be recovered. The acid was therefore carried on to
amide coupling.
(S)-4-(3-(benzyloxy)benzyloxy)-N-(1-cyanoethyl)benzamide (37)
[0304] General procedures D and E were used to deprotect 0.45 mmols
of (S)-tert-butyl 1-cyanoethylcarbamate and couple to 0.45 mmols of
the acid 36. After purification by flash chromatography, 0.23 mmols
of the title product was recovered. .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta. 7.78-7.72 (m, 2H), 7.47-7.42 (m, 2H), 7.41-7.37
(m, 2H), 7.36-7.29 (m, 2H), 7.06 (s, 1H), 7.02 (s, 1H), 7.01-6.98
(m, 2H), 6.95 (dd, J=2.5, 8.2, 1H), 6.46 (s, 1H), 5.15 (dd, J=7.6,
14.9, 1H), 5.09 (s, 2H), 5.07 (s, 2H), 1.65 (d, J=7.2, 3H).
.sup.13C NMR (75 MHz, CDCl.sub.3) .delta. 191.04, 163.88, 159.36,
137.82, 136.98, 132.24, 130.09, 128.86, 128.29, 127.74, 120.05,
115.39, 114.79, 114.14, 70.25.
VPC95219
[0305] General procedure H was used (with ammonium bromide in place
of ammonium chloride) to convert 0.22 mmols of 37 to the
corresponding amidine. Purification procedure B was used to purify
the title product, which was carried on for biological evaluation.
t.sub.R=9.5 min.; m/z=404.15.
4-(7-(cyclohexylmethoxy)heptyl)benzaldehyde (38)
[0306] General procedure M was used to convert 2.45 mmol of 11 and
2.70 mmol of 1-bromo-4-nitrobenzene to 1.89 mmol (77%) of the title
compound.
4-(7-(cyclohexylmethoxy)heptyl)benzoic acid (39)
[0307] General procedure I was used to convert 1.89 mmol of 38 to
1.89 mmol (100%) of the title compound.
(S)-1-(4-(7-(cyclohexylmethoxy)heptyl)benzoyl)pyrrolidine-2-carbonitrile
(40)
[0308] General procedure G was used to convert 2.77 mmols of 39 to
an acid chloride. After standard purification techniques, the
acyl-chloride was isolated in quantitative yield and immediately to
coupled 2.77 mmols of the TFA salt of(S)-pyrrolidine-2-carbonitrile
according to general procedure F. After standard purification
techniques, 1.23 mmols (44%) of the title product was
recovered.
VPC143237
[0309] General procedure H was used to convert 1.23 mmol of 21 to
0.780 mmol (84%) of the title product. The amidine HCl sat is a
clear gel that was isolated via flash chromatography with 15%
MeOH/CHCl.sub.3. The fractions were concentrated and the oil was
filtered through a fine frit with chloroform to remove traces of
silica gel. The clear and colorless oil was then mixed with 2N HCl
in Et.sub.2O and concentrated to the pure amidine HCl salt and
submitted for biological evaluation.
(R)-tert-butyl 2-cyanopyrrolidine-1-carboxylate (41)
[0310] To a solution of 2 grams (9.3 mmols) of N-Boc-D-Proline in
19 mL of dichloromethane was added 4.2 grams (42 mmols) of
triethylamine. The reaction mixture was cooled to -25.degree. C.
and 2.5 grams (18.6 mmols) of iso-butylchloroformate was added
dropwise. After stirring at this low temperature for 45 minutes,
0.83 grams (49 mmols) of ammonia was added to the mixture as a 7M
solution in methanol. The reaction vessel was warmed slowly to
ambient temperature and the solution was stirred for an additional
16 hours. After this time, the reaction mixture was evaporated to
dryness and reconstituted in 200 mL of EtOAc. The organic layer was
washed with four 15 mL portions of 1N HCl, dried with MgSO.sub.4
and evaporated to dryness. The crude material was immediately
subjected to general procedure E using 2 grams (18.9 mmols) of
triethylamine and 2.3 grams (11.16 mmols) of trifluoroacetic
anhydride in 93 mL of THF. After previously described purification
procedures (column chromatography, 25% EtOAc in hexanes), 0.8 grams
(4.1 mmols, 44%) of the title product was recovered. .sup.1H and
.sup.13C NMR data were identical to that reported for 28.
(R)-1-(4-dodecylbenzoyl)pyrrolidine-2-carbonitrile (42)
[0311] General procedure D was used to deprotect 1.4 mmols of 41
using 14 mL of dichloromethane and trifluoroacetic acid. After
previously described recrystallization techniques, the TFA salt of
41 was coupled to 0.4 grams (1.4 mmols) of 4-dodecylbenzoic acid
using 0.73 grams (1.4 mmols) of PyBOP and 0.72 grams (5.6 mmols) of
N,N-diisopropylethylamine in 14 mL of dichloromethane according to
general procedure E. Using previously described work-up and
purification techniques (column chromatography, 25% EtOAc in
hexanes), yielded 0.2 grams (0.55 mmols, 39%) of the title product
as an off-white solid. .sup.1H and .sup.13C NMR data were identical
to that of 29.
VPC96115
[0312] General procedure H was used to convert 0.27 grams (0.73
mmols) of 42 to the title product using 146 .mu.L of 0.5M sodium
methoxide in methanol (0.004 grams, 0.073 mmols), 7.3 mL of
anhydrous methanol and 0.043 grams (0.8 mmols) of ammonium
chloride. After previously described recrystallization procedures,
0.08 grams (0.19 mmols, 26%) of VPC96115 was recovered as a powdery
white solid and submitted for biological evaluation. .sup.1H and
.sup.13C NMR data were identical to that of VPC96091.
VPC96143
[0313] To a solution of 0.293 grams (0.83 mmols) of 29a in
anhydrous ethanol was added 0.132 grams (1.9 mmols) of
hydroxylamine hydrochloride and 0.202 grams (2 mmols) of
triethylamine. The mixture was heated to reflux for 2 hours at
which time it was cooled to ambient temperature. The mixture was
evaporated to dryness and taken up in 150 mL of chloroform. The
organic layer was washed with three 5 mL portions of deionized
water, one 5 mL portion of brine and dried of anhydrous MgSO.sub.4.
After purification by column chromatography (5% MeOH in
CHCl.sub.3), 0.83 mmols of the title product were recovered as an
off-white gelatinous solid. .sup.1H NMR (500 MHz, CDCl.sub.3)
.delta. 7.43 (d, J=7.9 Hz, 2H), 7.19 (d, J=7.8 Hz, 2H), 5.45 (s,
1H), 5.07-4.98 (m, 1H), 3.55 (t, J=5.5 Hz, 1H), 3.47 (s, 1H), 2.60
(t, J=7.6 Hz, 2H), 2.37 (d, J=5.7 Hz, 1H), 2.02 (dd, J=19.5, 10.0
Hz, 2H), 1.79 (dd, J=11.3, 5.9 Hz, 1H), 1.67-1.50 (m, 2H), 1.26 (m,
16H), 0.86 (t, J=6.9 Hz, 3H). .sup.13C NMR (126 MHz, CDCl.sub.3) ?
172.00, 154.17, 145.53, 133.46, 128.27, 127.32, 59.48, 56.67,
50.34, 35.81, 31.89, 31.23, 29.56, 29.47, 29.24, 26.69, 25.31,
22.67, 14.12.
[0314] The assays below are standard literature reported assays
known in the art for confirming and quantifying the activity of the
disclosed compounds.
Single Point Sphingosine Kinase Assay
[0315] Sphingosine kinase activity was assessed as described
previously (Kharel, Y., Lee, S., Snyder, A. H., Sheasley-O'Neill,
S. L., Morris, M. A., Setiady, Y., Zhu, R, Zigler, M. A., Burcin,
T. L., Ley, K., Tung, K. S. K., Engelhard, V. H., Macdonald, T. L.
and Lynch, K. R. Sphingosine kinase 2 is required for modulation of
lymphocyte traffic by FTY720. J Biological Chemistry 280:
36865-36872 (2005)). High levels of human sphingosine kinase type 1
(mSK1) and mouse sphingosine kinase type 2 (mSK2) were expressed in
Sf9 insect cells by infection with cognate baculoviruses. Crude
homogenates were incubated with .alpha.-[.sup.32P]ATP and 10
micromolar D-erythro-sphingosine in the presence of 100 micromolar
concentrations of test compounds for 20 minutes at 37 C. The
product, radiolabeled S1P, was isolated by extraction into organic
solvents after acidification and displaced by thin layer
chromatography. The radiolabeled S1P was scraped using the
autoradiogram as a template and the amount of material determine by
liquid scintillation counting. The results are illustrated in Table
1.
Determination of Ki Values.
[0316] Crude homogenates of Sf9 cells containing high levels of
recombinant mouse SphK2 or human SphK1 were incubated with
.gamma.-[.sup.32P]ATP and various concentrations (0 to 500
micromolar) of D-erythro-sphingosine for 20 minutes hour at 37 C in
the presence of fixed concentration of test compound. The product,
radiolabeled S1P, was isolated by extraction into organic solvents
followed by thin layer chromatography, recovered and radioactivity
measured by liquid scintillation counting. The Ki values so
determined are given in Table 1 (above).
Exemplification of Pro-Drug Activity
[0317] Mice were dosed with 20 mpk of amidoxime, VPC96143, both IP
and orally. S1P in plasma and organs was measured at times ASAP, 2
hrs and 24 hrs. The compound amidine, VPC96077, was also measured
in plasma and organs at times ASAP, 2 hrs and 24 hrs.
[0318] Mice Numbering [0319] 51: dosed with 20 mpk amidoxime
VPC96143 administered IP. Plasma extracted ASAP and 2 hrs. Organs
harvested at 2 hrs. [0320] 52: dosed with 20 mpk amidoxime VPC96143
administered IP. Plasma extracted ASAP and 24 hrs. Organs harvested
at 2 hrs. [0321] 53: dosed with 20 mpk amidoxime VPC96143
administered orally. Plasma extracted ASAP and 2 hrs. Organs
harvested at 2 hrs.
[0322] The results are illustrated in FIGS. 7A-7D.
[0323] U937 cells (5 million) were treated with 20 .mu.M of
amidoxime, VPC96143. The accumulation of S1P was measured at 1 hr,
4 hr and 24 hrs. The levels of amidine, VPC96077 were also measured
at 1 hr, 4 hrs and 24 hrs. The results are illustrated in FIGS.
8A-8B.
[0324] U937 cells (5 million) were treated with amidoxime,
VPC96143, at concentrations of 1 .mu.M or 10 .mu.M, and amidine,
VPC96077, at concentrations of 1 .mu.M or 10 .mu.M. The
accumulation of S1P was measured 2 hrs post-dose. Also, the levels
of amidine, VPC96077, were measured 2 hrs post-dose. The results
are illustrated in FIGS. 9A-9B.
[0325] The accumulation of S1P and amidine and the drug to pro-drug
ratio in organs, demonstrates the efficiency of conversion of
amidoxime to amidine in target organs indicating that amidoximes
are pro-drug forms of amidines.
[0326] The invention should not be construed to be, limited solely
to the assays and methods described above, but should be construed
to include other methods and assays as well. Other methods that are
used but not described above are well known and within the
competence of one of ordinary skill in the art of chemistry,
biochemistry, molecular biology, and clinical medicine. One of
ordinary skill in the art will know that other assays and methods
are available to perform the procedures described above.
[0327] The abbreviations used above have their conventional meaning
within the clinical, chemical, and biological arts. In the case of
any inconsistencies, the present disclosure, including any
definitions therein will prevail.
* * * * *